Organic matrices in metazoan calcium carbonate skeletons: composition, functions, evolution.

2016

9 pages; International audience; Calcium carbonate skeletal tissues in metazoans comprise a small quantity of occluded organic macromolecules, mostly proteins and polysaccharides that constitute the skeletal matrix. Because its functions in modulating the biomineralization process are well known, the skeletal matrix has been extensively studied, successively via classical biochemical approaches, via molecular biology and, in recent years, via transcriptomics and proteomics. The optimistic view that the deposition of calcium carbonate minerals requires a limited number of macromolecules has been challenged, in the last decade, by high-throughput approaches. Such approaches have made possible…

"Save Private Pinna !" The Mediterranean fan mussel, a patrimonial bivalve, is in serious danger of extinction.

2019

Pinna nobilis is the largest bivalve in the Mediterranean Sea. This endemic species lives in the Posidonia meadows,in the form of small sparse populations. It is a heritage species: its byssus has long been collected to be wovenand used to make gloves, hats or stoles. Protected by a European Directive (1992), P. nobilis is closely monitored,nationally and internationally. But since the end of 2016, an epidemic, which origin seems to be a protozoan parasite,decimates populations (up to 100% mortality). This event began on the Spanish coast and spreads now all around theMediterranean. Although conservation measures have been taken, the next few years could see the extinction of thisemblematic…

The Role of Enzymes in Biomineralization Processes

2010

Molluscan shell proteins

2004

Abstract The shell secreted by molluscs is one of the most remarkable examples of a matrix-mediated mineralisation performed outside living tissues. The calcifying matrix is a mixture of proteins, glycoproteins, and polysaccharides that precisely self-assemble and control the CaCO 3 polymorph (calcite, aragonite), the size, the shapes of the crystallites, and finally, the texture of the shell. In spite of several biochemical studies, the molecular aspects of the shell building are far from being understood. The present article makes an overview of the most recent molecular data on the proteinaceous components of the shell matrix. These data put into question the classical models of mollusca…

Characterization of MRNP34, a novel methionine-rich nacre protein from the pearl oysters

2012

9 pages; International audience; Nacre of the Pinctada pearl oyster shells is composed of 98% CaCO(3) and 2% organic matrix. The relationship between the organic matrix and the mechanism of nacre formation currently constitutes the main focus regarding the biomineralization process. In this study, we isolated a new nacre matrix protein in P. margaritifera and P. maxima, we called Pmarg- and Pmax-MRNP34 (methionine-rich nacre protein). MRNP34 is a secreted hydrophobic protein, which is remarkably rich in methionine, and which is specifically localised in mineralizing the epithelium cells of the mantle and in the nacre matrix. The structure of this protein is drastically different from those …

Deep conservation of bivalve nacre proteins highlighted by shell matrix proteomics of the Unionoida Elliptio complanata and Villosa lienosa.

2016

The formation of the molluscan shell nacre is regulated to a large extent by a matrix of extracellular macromolecules that are secreted by the shell-forming tissue, the mantle. This so-called ‘calcifying matrix’ is a complex mixture of proteins, glycoproteins and polysaccharides that is assembled and occluded within the mineral phase during the calcification process. Better molecular-level characterization of the substances that regulate nacre formation is still required. Notable advances in expressed tag sequencing of freshwater mussels, such as Elliptio complanata and Villosa lienosa , provide a pre-requisite to further characterize bivalve nacre proteins by a proteomic approach. In this…

Thermal stability of nacre proteins of the polynesian pearl oyster: a proteomic study.

2015

Mollusc shells are organic-inorganic composites that are often preserved in the fossil record. However, the way the organic fraction, also called shell matrix, gets fossilized remains an unsolved question, in spite of several old and more recent studies. In the present paper, we have tried to mimic a diagenetic process by constantly heating for ten days at 100°C fresh nacre powder samples of the Polynesian pearl oyster Pinctadamargaritifera. Each day, aliquots of nacre powder were sampled and the matrix was subsequently extracted. It was further analysed by direct weigh quantification, by immunological techniques and by proteomics. Our preliminary data suggest that nacre proteins, when heat…

The skeletal proteome of the coral Acropora millepora: the evolution of calcification by co-option and domain shuffling.

2013

14 pages; International audience; In corals, biocalcification is a major function that may be drastically affected by ocean acidification (OA). Scleractinian corals grow by building up aragonitic exoskeletons that provide support and protection for soft tissues. Although this process has been extensively studied, the molecular basis of biocalcification is poorly understood. Notably lacking is a comprehensive catalog of the skeleton-occluded proteins-the skeletal organic matrix proteins (SOMPs) that are thought to regulate the mineral deposition. Using a combination of proteomics and transcriptomics, we report the first survey of such proteins in the staghorn coral Acropora millepora. The or…

The skeleton of the staghorn coral Acropora millepora: molecular and structural characterization.

2014

15 pages; International audience; The scleractinian coral Acropora millepora is one of the most studied species from the Great Barrier Reef. This species has been used to understand evolutionary, immune and developmental processes in cnidarians. It has also been subject of several ecological studies in order to elucidate reef responses to environmental changes such as temperature rise and ocean acidification (OA). In these contexts, several nucleic acid resources were made available. When combined to a recent proteomic analysis of the coral skeletal organic matrix (SOM), they enabled the identification of several skeletal matrix proteins, making A. millepora into an emerging model for biomi…

Shell repair in the clam Ruditapes philippinarum, affected by the Brown Ring Disease (BRD): a biochemical and serological study.

2009

AbstractFor more than two decades, the edible clam Ruditapes philippinarum has been affected by the Brown Ring Disease (BRD), a bacterial infection characterized by the formation of a brown organic deposit in the internal side of the valves. Although this infection is often lethal, in some cases specimens can overcome it by remineralizing over the organic deposit. The goal of the present study is to compare biochemically and immunologically the shell matrices of repaired and healthy zones. Our data suggest that the repair zones exhibit certain variability, which would be the direct consequence of a modification of the secretory regime of calcifying tissues responsible of the repair process.

The shell organic matrix of the crossed lamellar queen conch shell (Strombus gigas)

2014

10 pages; International audience; In molluscs, the shell organic matrix comprises a large set of biomineral-occluded proteins, glycoproteins and polysaccharides that are secreted by the calcifying mantle epithelium, and are supposed to display several functions related to the synthesis of the shell. In the present paper, we have characterized biochemically the shell matrix associated to the crossed-lamellar structure of the giant queen conch Strombus gigas. The acid-soluble (ASM) and acid-insoluble (AIM) matrices represent an extremely minor fraction of the shell. Both are constituted of polydisperse and of few discrete proteins among which three fractions, obtained by preparative SDS-PAGE …

Differences between Bond Lengths in Biogenic and Geological Calcite

2010

8 pages; International audience; We used high-resolution neutron powder diffraction to accurately measure the atomic positions and bond lengths in biogenic and geological calcite. A special procedure for data analysis was developed in order to take into account the considerable amounts of magnesium present in all the investigated samples. As a result, in biogenic calcite we found some atomic bonds to have significantly different lengths as compared to those in geological calcite, after the contribution of magnesium is accounted for. The maximum effect (elongation up to 0.7%) was found for the C−O bonds. We also analyzed changes in frequencies and spectral widths of normal vibrations of carb…

Carbonic anhydrase and metazoan biocalcification: a focus on molluscs.

2015

Carbonic anhydrase is a super-family of metallo-enzymes (containing α, β, γ, ζ and δ-CA families) that catalyse the reversible hydration of carbon dioxide. Among their numerous functions, CAs - in particular that of the α-CA family - are known to play a key role in biocalcification processes, i.e., the ability to deposit calcium carbonate crystallites in a controlled manner to form exoskeletons. In the gastropod mollusc Haliotistuberculata – the European abalone – we identified two CA transcripts, htCA1 and htCA2, in the mantle, the calcifying organ responsible for shell formation from an extracellular organic matrix and a mixture of inorganic ions. Because these two transcripts are specifi…

The shell-forming proteome of Lottia gigantea reveals both deep conservations and lineage-specific novelties

2013

19 pages; International audience; Proteins that are occluded within the molluscan shell, the so-called shell matrix proteins (SMPs), are an assemblage of biomolecules attractive to study for several reasons. They increase the fracture resistance of the shell by several orders of magnitude, determine the polymorph of CaCO(3) deposited, and regulate crystal nucleation, growth initiation and termination. In addition, they are thought to control the shell microstructures. Understanding how these proteins have evolved is also likely to provide deep insight into events that supported the diversification and expansion of metazoan life during the Cambrian radiation 543 million years ago. Here, we p…

Variability of shell repair in the Manila clam Ruditapes philippinarum affected by the Brown Ring Disease: A microstructural and biochemical study

2011

11 pages; International audience; For more than two decades, the Manila clam Ruditapes philippinarum has been regularly affected by Brown Ring Disease (BRD), an epizootic event caused by the bacterium Vibrio tapetis and characterized by the development of a brown deposit on the inner face of valves. Although BRD infection is often lethal, some clams recover by mineralizing a new repair shell layer, which covers the brown deposit and fully isolates it from living tissues. In order to understand this specific shell repair process, the microstructures of repaired zones were compared to those of shells unaffected by BRD. In addition, the organic matrix associated with unaffected shells and to r…

Smaller calcite lattice deformation caused by occluded organic material in coccoliths than in mollusk shell.

2015

7 pages; International audience; The growth and nucleation of biominerals are directed and affected by associated biological molecules. In this paper, we investigate the influence of occluded biomolecules on biogenic calcite from the coccolithophorid Pleurochrysis carterae and from chalk, a rock composed predominantly of fossil coccoliths. We compare the results with data on chalk from the extensively studied mussel Pinna nobilis that served as a control. Using high resolution synchrotron powder X-ray diffraction combined with in situ heating, the influence of organic compounds on the structure of the inorganic phase was probed. Two heating cycles allow us to differentiate the effects of th…

Pmarg-pearlin is a matrix protein involved in nacre framework formation in the pearl oyster Pinctada margaritifera.

2011

11 pages; International audience; The shell of pearl oysters is organized in multiple layers of CaCO(3) crystallites packed together in an organic matrix. Relationships between the components of the organic matrix and mechanisms of nacre formation currently constitute the main focus of research into biomineralization. In this study, we characterized the pearlin protein from the oyster Pinctada margaritifera (Pmarg); this shares structural features with other members of a matrix protein family, N14/N16/pearlin. Pmarg pearlin exhibits calcium- and chitin-binding properties. Pmarg pearlin transcripts are distinctively localized in the mineralizing tissue responsible for nacre formation. More s…

Spine and test skeletal matrices of the Mediterranean sea urchinArbacia lixula- a comparative characterization of their sugar signature

2015

15 pages; International audience; Calcified structures of sea urchins are biocomposite materials that comprise a minor fraction of organic macromolecules, such as proteins, glycoproteins and polysaccharides. These macromolecules are thought to collectively regulate mineral deposition during the process of calcification. When occluded, they modify the properties of the mineral. In the present study, the organic matrices (both soluble and insoluble in acetic acid) of spines and tests from the Mediterranean black sea urchin Arbacia lixula were extracted and characterized, in order to determine whether they exhibit similar biochemical signatures. Bulk characterizations were performed by mono-di…

Phosphorylation of serine residues is fundamental for the calcium-binding ability of Orchestin, a soluble matrix protein from crustacean calcium stor…

2003

International audience; Orchestia cavimana is a terrestrial crustacean, which cyclically stores calcium in diverticula of the midgut, in the form of calcified amorphous concretions. These concretions are associated with a proteinaceous matrix, the main constituent of the soluble matrix is Orchestin, an acidic calcium-binding protein [Testenière et al., Biochem. J. 361 (2002) 327-335]. In the present paper, we clearly demonstrate that Orchestin is phosphorylated on serine and tyrosine residues, but that calcium binding only occurs via the phosphoserine residues. To our knowledge, this is the first example of an invertebrate mineralization for which a post-translational modification is clearl…

Anisotropic lattice distortions in biogenic calcite induced by intra-crystalline organic molecules.

2006

9 pages; International audience; We have performed precise structural measurements on five different calcitic seashells by high-resolution X-ray powder diffraction on a synchrotron beam line and by laboratory single crystal X-ray diffraction. The unit cell parameters a and c of biogenic calcite were found to be systematically larger than those measured in the non-biogenic calcite. The maximum lattice distortion (about 2.10(-3)) was detected along the c-axis. Under heat treatment above 200 degrees C, a pronounced lattice relaxation was observed, which allowed us to conclude that anisotropic lattice swelling in biogenic calcite is induced by organic macromolecules incorporated within the sing…

Characterization of molecular processes involved in the pearl formation in Pinctada margaritifera for the sustainable development of pearl farming in…

2011

11 pages; International audience

The shell of the invasive bivalve species Dreissena polymorpha: biochemical, elemental and textural Investigations.

2016

28 pages; International audience; The zebra mussel Dreissena polymorpha is a well-established invasive model organism. Although extensively used in environmental sciences, virtually nothing is known of the molecular process of its shell calcification. By describing the microstructure, geochemistry and biochemistry/proteomics of the shell, the present study aims at promoting this species as a model organism in biomineralization studies, in order to establish a bridge with ecotoxicology, while sketching evolutionary conclusions. The shell of D. polymorpha exhibits the classical crossed-lamellar/complex crossed lamellar combination found in several heterodont bivalves, in addition to an extern…

Biochemical Characterization of the Soluble Organic Matrix of Gastroliths from Decapods.

2005

10 pages; International audience; Gastroliths are disc-shaped calcium storage structures cyclically elaborated by some decapods in their stomach wall. In the present study, we extracted the gastrolith organic matrix from three different decapods, two crayfishes and one lobster, and began to characterize the soluble matrix obtained after decalcification by acetic acid. By using different staining (silver nitrate, Coomassie Blue, Stains-all) after SDS-PAGE, we evidenced that these soluble fractions are rich in polypeptides, some seems common, others are completely different. Among them, putative calcium binding proteins might be present and have to be further characterized. This soluble organ…

Biomineralisations in crustaceans: storage strategies

2004

Abstract Crustaceans are a remarkable group of animals because of their ability to elaborate cyclically two kinds of calcified biomineralisations: an unstretchable exoskeleton (or cuticle) and also, for many species, depending of the way of life of the considered animal, transitory calcium deposits. They are in consequence subjected to a cyclic life and notably, to a periodical balance between two sources of calcium, exogenous and/or endogenous. The storage structures, essentially composed of calcium carbonate precipitated within a proteinaceous organic matrix, are very diversified. The calcium carbonate of the cuticle of most of the species is under a crystalline state and/or an amorphous …

The ‘Shellome’ of the Crocus Clam Tridacna crocea Emphasizes Essential Components of Mollusk Shell Biomineralization

2021

Molluscan shells are among the most fascinating research objects because of their diverse morphologies and textures. The formation of these delicate biomineralized structures is a matrix-mediated process. A question that arises is what are the essential components required to build these exoskeletons. In order to understand the molecular mechanisms of molluscan shell formation, it is crucial to identify organic macromolecules in different shells from diverse taxa. In the case of bivalves, however, taxon sampling in previous shell proteomics studies are focused predominantly on representatives of the class Pteriomorphia such as pearl oysters, edible oysters and mussels. In this study, we hav…

Climate variation during the Holocene influenced the skeletal properties of Chamelea gallina shells in the North Adriatic Sea (Italy)

2021

Understanding how marine taxa will respond to near-future climate changes is one of the main challenges for management of coastal ecosystem services. Ecological studies that investigate relationships between the environment and shell properties of commercially important marine species are commonly restricted to latitudinal gradients or small-scale laboratory experiments. This paper aimed to explore the variations in shell features and growth of the edible bivalve Chamelea gallina from the Holocene sedimentary succession to present-day thanatocoenosis of the Po Plain-Adriatic Sea system (Italy). Comparing the Holocene sub-fossil record to modern thanatocoenoses allowed obtaining an insight o…

The shell matrix of the european thorny oyster, Spondylus gaederopus: microstructural and molecular characterization.

2020

17 pages; International audience; Molluscs, the largest marine phylum, display extraordinary shell diversity and sophisticated biomineral architectures. However, mineral-associated biomolecules involved in biomineralization are still poorly characterised.We report the first comprehensive structural and biomolecular study of Spondylus gaederopus, a pectinoid bivalve with a peculiar shell texture. Used since prehistoric times, this is the best-known shell of Europe’s cultural heritage. We find that Spondylus microstructure is very poor in mineral-bound organics, which are mostly intercrystalline and concentrated at the interface between structural layers.Using high-resolution liquid chromatog…

Protein mapping of calcium carbonate biominerals by immunogold

2007

The construction of metazoan calcium carbonate skeletons is finely regulated by a proteinaceous extracellular matrix, which remains embedded within the exoskeleton. In spite of numerous biochemical studies, the precise localization of skeletal proteins has remained for a long time as an elusive goal. In this paper, we describe a technique for visualizing shell matrix proteins on the surface of calcium carbonate crystals or within the biominerals. The technique is as follows: freshly broken pieces of biominerals or NaOCl then EDTA-etched polished surfaces are incubated with an antibody elicited against one matrix protein, then with a secondary gold-coupled antibody. After silver enhancement,…

Ultrastructure of the Interlamellar Membranes of the Nacre of the Bivalve Pteria hirundo, Determined by Immunolabelling.

2015

The current model for the ultrastructure of the interlamellar membranes of molluscan nacre imply that they consist of a core of aligned chitin fibers surrounded on both sides by acidic proteins. This model was based on observations taken on previously demineralized shells, where the original structure had disappeared. Despite other earlier claims, no direct observations exist in which the different components can be unequivocally discriminated. We have applied different labeling protocols on non-demineralized nacreous shells of the bivalve Pteria. With this method, we have revealed the disposition and nature of the different fibers of the interlamellar membranes that can be observed on the …

Data mining approaches to identify biomineralization related sequences.

2015

Proteomics is an efficient high throughput technique developed to identify proteins from a crude extract using sequence homology. Advances in Next Generation Sequencing (NGS) have led to increase knowledge of several non-model species. In the field of calcium carbonate biomineralization, the paucity of available sequences (such as the ones of mollusc shells) is still a bottleneck in most proteomic studies. Indeed, this technique needs proteins databases to find homology. The aim of this study was to perform different data mining approaches in order to identify novel shell proteins. To this end, we disposed of several publicly non-model molluscs databases. Previously identified molluscan she…

Pearl grafting: tracking the biological origin of nuclei by straightforward immunological methods.

2018

9 pages; International audience; French Polynesia is renowned for the production of Tahitian black pearl. These gems are obtained by grafting a nucleus into the gonad of a receiving oyster together with a graft, i.e. a small section of mantle tissue of a donor oyster. This procedure initiates the formation of a pearl sack around the nucleus, and subsequently, the deposition of concentric layers of nacre. The nucleus plays a key-role in pearl formation and its characteristics influence markedly the quality of the final product. As it is manufactured from mollusc shells, it contains a small percentage of organics. In the present paper, we used a set of biochemical techniques to characterize a…

Nacre evolution: a proteomic approach.

2009

AbstractFrom an evolutionary viewpoint, the molluscan nacre constitutes a fascinating object. This microstructure appeared early, in the Lower Cambrian period, about 530 million years ago, and since then, has been kept unchanged until today. Nacre is restricted to the conchiferan mollusks, where it occurs in t least three main classes, bivalves, gastropods and cephalopods. The aim of the present study is to investigate whether all nacres are built from the same “macromolecular tools”, proteins of the nacre matrix. To this end, we studied three new nacre models, the freshwater bivalve Unio pictorum, the cephalopod Nautilus macromphalus, and the gastropod Haliotis asinina, to which we applied…

Proteomics of CaCO3 biomineral-associated proteins: how to properly address their analysis.

2013

8 pages; International audience; In a recent editorial (Proc. Natl. Acad. Sci., 2013 110, E2144-E2146) and elsewhere, questions have been raised regarding the experimental practices in relation to the proteomic analysis of organic matrices associated to the biomineralized CaCO3 skeletons of metazoans such as molluscan shells and coral skeletons. Indeed, although the use of new high sensitivity MS technology potentially allows to identify a greater number of proteins, it is also equally (or even more) sensitive to contamination of residual proteins from soft tissues, which are in close contact with the biomineral. Based on our own past and present experimental know-how-observations that are …

Molluscan Shell Proteins: Primary Structure, Origin, and Evolution

2007

In the last few years, the field of molluscan biomineralization has known a tremendous mutation, regarding fundamental concepts on biomineralization regulation as well as regarding the methods of investigation. The most recent advances deal more particularly with the structure of shell biominerals at nanoscale and the identification of an increasing number of shell matrix protein components. Although the matrix is quantitatively a minor constituent in the shell of mollusks (less than 5% w/w), it is, however, the major component that controls different aspects of the shell formation processes: synthesis of transient amorphous minerals and evolution to crystalline phases, choice of the calciu…

Biochemical characterization of the skeletal matrix of the massive coral, Porites australiensis - The saccharide moieties and their localization.

2018

11 pages; International audience; To construct calcium carbonate skeletons of sophisticated architecture, scleractinian corals secrete an extracellular skeletal organic matrix (SOM) from aboral ectodermal cells. The SOM, which is composed of proteins, saccharides, and lipids, performs functions critical for skeleton formation. Even though polysaccharides constitute the major component of the SOM, its contribution to coral skeleton formation is poorly understood. To this end, we analyzed the SOM of the massive colonial coral, Porites australiensis, the skeleton of which has drawn great research interest because it records environmental conditions throughout the life of the colony. The coral …

Nanoscale assembly processes revealed in the nacroprismatic transition zone of Pinna nobilis mollusc shells

2015

Intricate biomineralization processes in molluscs engineer hierarchical structures with meso-, nano-, and atomic architectures that give the final composite material exceptional mechanical strength and optical iridescence on the macroscale. This multiscale biological assembly inspires new synthetic routes to complex materials. Our investigation of the prism-nacre interface reveals nanoscale details governing the onset of nacre formation using high-resolution scanning transmission electron microscopy. A wedge polishing technique provides unprecedented, large-area specimens required to span the entire interface. Within this region, we find a transition from nanofibrillar aggregation to irregu…

Characterization of the teeth skeletal matrix from Arbacia lixula.

2015

15 pages; International audience

Caspartin: thermal stability and occurrence in mollusk calcified tissues.

2005

8 pages; International audience; The fan mussel Pinna nobilis secretes a two-layered shell, the external layer of which is composed of long calcitic prisms. The dissolution of these prisms releases an assemblage of acidic intracrystalline proteins, among which caspartin, an Asp-rich protein (Marin et al., 2005). A polyclonal antibody raised against purified caspartin was used for estimating its thermal stability, and for investigating the presence of cross-reacting proteins in the shell matrices of several mollusks. In the first case, prisms of Pinna nobilis were heated at 100°C, for one to eleven days. The degradation of caspartin was followed on SDS-PAGE and on Western-blot. The experimen…

Single nanogranules preserve intracrystalline amorphicity in biominerals.

2015

We revisit the ultrastructural features of different calcareous biominerals and identify remarkable similarities: taxonomically very distant species show a common nanogranular structure, even if different extracellular secretion patterns are employed or calcium carbonate polymorphs formed. By these analyses, we elucidate the locus of the small fraction of intracrystalline organic matrix revealing its intergranular character and localize the intracrystalline amorphous calcium carbonate moiety commonly found in mesocrystalline biominerals and provide a first explanation for the pathway by which it is preserved.

A new twist on sea silk : the peculiar protein ultrastructure of fan shell and pearl oyster byssus

2018

11 pages; International audience; Numerous mussel species produce byssal threads - tough proteinaceous fibers, which anchor mussels in aquatic habitats. Byssal threads from Mytilus species, which are comprised of modified collagen proteins - have become a veritable archetype for bio-inspired polymers due to their self-healing properties. However, threads from different species are comparatively much less understood. In particular, the byssus of Pinna nobilis comprises thousands of fine fibers utilized by humans for millennia to fashion lightweight golden fabrics known as sea silk. P. nobilis is very different from Mytilus from an ecological, morphological and evolutionary point of view and …

Heavy Metals in Mollusc Shells: A Quick Method for their Detection

2015

It is known for a long time that calcified tissues secreted by aquatic or terrestrial invertebrates – such as mollusc shells – have the ability to concentrate large amounts of pollutants, in particular heavy metals. In the present paper, we have found an extremely rapid and easy procedure to qualitatively detect the putative presence of heavy metals in shells, without having to use sophisticated techniques such as Wavelength Dispersive Spectroscopy, atomic adsorption spectroscopy or ICP-MS. Our method rests on the capacity of the silver enhancement chemicals that are traditionally used in immunogold localization experiments to increase the size of heavy metal nanoparticles, whatever the che…

Biomineralix (COST action TD0903), 2009-2014: an overview.

2015

Biomineralization is the process by which living systems produce minerals. At theEuropean scale, this particular scientific field associates researchers of diverse horizons,ranging from zoology, mineralogy, palaeontology, and environmental sciences to materialssciences, dentistry and bone surgery. In this context, the COST Action TD0903, also referredas Biomineralix, was set up in November 2009 under the auspices of the COST Office inBrussels. This scientific network was active during four and half years, until May 2014. Itassociated 17 European countries, and a total of 29 laboratories. Its main objectives weremultiple: 1. To promote biochemical and proteomic research on the biomineralisat…

Genesis of amorphous calcium carbonate containing alveolar plates in the ciliate Coleps hirtus (Ciliophora, Prostomatea).

2013

7 pages; International audience; In the protist world, the ciliate Coleps hirtus (phylum Ciliophora, class Prostomatea) synthesizes a peculiar biomineralized test made of alveolar plates, structures located within alveolar vesicles at the cell cortex. Alveolar plates are arranged by overlapping like an armor and they are thought to protect and/or stiffen the cell. Although their morphology is species-specific and of complex architecture, so far almost nothing is known about their genesis, their structure and their elemental and mineral composition. We investigated the genesis of new alveolar plates after cell division and examined cells and isolated alveolar plates by electron microscopy, e…

Nautilin-63, a novel acidic glycoprotein from the shell nacre of Nautilus macromphalus

2011

In molluscs, and more generally in metazoan organisms, the production of a calcified skeleton is a complex molecular process that is regulated by the secretion of an extracellular organic matrix. This matrix constitutes a cohesive and functional macromolecular assemblage, containing mainly proteins, glycoproteins and polysaccharides that, together, control the biomineral formation. These macromolecules interact with the extruded precursor mineral ions, mainly calcium and bicarbonate, to form complex organo-mineral composites of well-defined microstructures. For several reasons related to its remarkable mechanical properties and to its high value in jewelry, nacre is by far the most studied …

Shell palaeoproteomics: first application of peptide mass fingerprinting for the rapid identification of mollusc shells in archaeology.

2020

10 pages; International audience; Molluscs were one of the most widely-used natural resources in the past, and their shells are abundant among archaeological findings. However, our knowledge of the variety of shells that were circulating in prehistoric times (and thus their socio-economic and cultural value) is scarce due to the difficulty of achieving taxonomic determination of fragmented and/or worked remains. This study aims to obtain molecular barcodes based on peptide mass fingerprints (PMFs) of intracrystalline proteins, in order to obtain shell identification. Palaeoproteomic applications on shells are challenging, due to low concentration of molluscan proteins and an incomplete unde…

Molecular modularity and asymmetry of the molluscan mantle revealed by a gene expression atlas

2018

15 pages; International audience; Background: Conchiferan molluscs construct a biocalcified shell that likely supported much of their evolutionary success.However, beyond broad proteomic and transcriptomic surveys of molluscan shells and the shell-forming mantle tissue,little is known of the spatial and ontogenetic regulation of shell fabrication. In addition, most efforts have been focused onspecies that deposit nacre, which is at odds with the majority of conchiferan species that fabricate shells using acrossed-lamellar microstructure, sensu lato. Results: By combining proteomic and transcriptomic sequencing with in situhybridization we have identified a suite of gene products associated …

Mineral phase in shell repair of Manila clam Venerupis philippinarum affected by brown ring disease.

2011

14 pages; International audience; The mineral phase of shell repair in the Manila clam Venerupis philippinarum affected by brown ring disease (BRD) was characterised at various scales and at various stages of shell repair by confocal Raman microspectrometry and scanning electron microscopy. Spherulitic and quadrangular aragonite microstructures associated with polyene pigments were clearly observed. Von Kossa staining showed that at the beginning of shell repair, hemocytes are filled with insoluble calcium carbonate salts in all fluids and then are transported toward the extrapallial fluids and the repair sites. Our analyses suggest that after a Vibrio tapetis attack and BRD deposit some cl…

The test skeletal matrix of the black sea urchin Arbacia lixula

2015

11 pages; International audience; In the field of biomineralization, the past decade has been marked by the increasing use of high throughput techniques, i.e. proteomics, for identifying in one shot the protein content of complex macromolecular mixtures extracted from mineralized tissues. Although crowned with success, this approach has been restricted so far to a limited set of key-organisms, such as the purple sea urchin Strongylocentrotus purpuratus, the pearl oyster or the abalone, leaving in the shadow non-model organisms. As a consequence, it is still unknown to what extent the calcifying repertoire varies, from group to group, at high (phylum, class), median (order, family) or low (g…

A minimal molecular toolkit for mineral deposition? Biochemistry and proteomics of the test matrix of adult specimens of the sea urchin Paracentrotus…

2016

12 pages; International audience; The sea urchin endoskeleton consists of a magnesium-rich biocalcite comprising a small amount of occluded organic macromolecules. This structure constitutes a key-model for understanding the mineral - organics interplay, and for conceiving in vitro bio-inspired materials with tailored properties. Here we employed a deep-clean technique to purify the occluded proteins from adult Paracentrotus lividus tests. We characterized them by 1- and 2D-electrophoreses, ELISA and immunoblotting, and using liquid chromatography coupled with Mass Spectrometry (nanoLC-MS/MS), we identified two metalloenzymes (carbonic anhydrase and MMP), a set of MSP130 family members, sev…

The shell matrix and microstructure of the Ram's Horn squid: Molecular and structural characterization.

2020

20 pages; International audience; Molluscs are one of the most diversified phyla among metazoans. Most of them produce an external calcified shell, resulting from the secretory activity of a specialized epithelium of the calcifying mantle. This biomineralization process is controlled by a set of extracellular macromolecules, the organic matrix. In spite of several studies, these components are mainly known for bivalves and gastropods. In the present study, we investigated the physical and biochemical properties of the internal planispiral shell of the Ram's Horn squid Spirula spirula. Scanning Electron Microscope investigations of the shell reveal a complex microstructural organization. The…

Structural commonalities and deviations in the hierarchical organization of crossed-lamellar shells: A case study on the shell of the bivalve Glycyme…

2016

11 pages; International audience; The structural organization of the palliostracum—the dominant part of the shell which is formed by the mantle cells—of Glycymeris glycymeris (Linné 1758) is comprised of five hierarchical levels with pronounced structural commonalities and deviations from other crossed-lamellar shells. The hierarchical level known as second order lamellae, present within other crossed-lamellar shells, is absent highlighting a short-coming of the currently used nomenclature. On the mesoscale, secondary microtubules penetrate the palliostracum and serve as crack arrestors. Moreover, the growth lamellae follow bent trajectories possibly impacting crack propagation, crack defle…

The degradation of intracrystalline mollusc shell proteins: a proteomics study of Spondylus gaederopus.

2021

Mollusc shells represent excellent systems for the preservation and retrieval of genuine biomolecules from archaeological or palaeontological samples. As a consequence, the post-mortem breakdown of intracrystalline mollusc shell proteins has been extensively investigated, particularly with regard to its potential use as a "molecular clock" for geochronological applications. But despite seventy years of ancient protein research, the fundamental aspects of diagenesis-induced changes to protein structures and sequences remain elusive. In this study we investigate the degradation of intracrystalline proteins by performing artificial degradation experiments on the shell of the thorny oyster, Spo…

'Palaeoshellomics' reveals the use of freshwater mother-of-pearl in prehistory

2019

The extensive use of mollusc shell as a versatile raw material is testament to its importance in prehistoric times. The consistent choice of certain species for different purposes, including the making of ornaments, is a direct representation of how humans viewed and exploited their environment. The necessary taxonomic information, however, is often impossible to obtain from objects that are small, heavily worked or degraded. Here we propose a novel biogeochemical approach to track the biological origin of prehistoric mollusc shell. We conducted an in-depth study of archaeological ornaments using microstructural, geochemical and biomolecular analyses, including ‘palaeoshellomics’, the first…

Biomineralization toolkit: the importance of sample cleaning prior to the characterization of biomineral proteomes.

2013

In an interesting work published recently in PNAS, Drake et al. (1) presented a proteomic study of the skeleton from the stony coral Stylophora pistillata . This study identified proteins that are associated to the mineral phase (i.e., that potentially contribute to shape the skeleton). In other words, this set of proteins is supposed to represent the so-called “biomineralization toolkit.” Although some of the 36 proteins reported in Drake et al. (1) appear as genuine extracellular matrix (ECM) proteins related to biomineralization, such as coral acid-rich proteins or carbonic anhydrase, some others are obvious intracellular contaminants that should not be considered as skeletal organic mat…

Nacre calcification in the freshwater mussel Unio pictorum: carbonic anhydrase activity and purification of a 95 kDa calcium-binding glycoprotein.

2008

9 pages; International audience; The formation of the molluscan shell is finely tuned by macromolecules of the shell organic matrix. Previous results have shown that the acid-soluble fraction of the nacre matrix of the freshwater paleoheterodont bivalve Unio pictorum shell displays a number of remarkable properties, such as calcium-binding activity, the presence of extensive glycosylations and the capacity to interfere at low concentration with in vitro calcium carbonate precipitation. Here we have found that the nacre-soluble matrix exhibits a carbonic anhydrase activity, an important function in calcification processes. This matrix is composed of three main proteinaceous discrete fraction…

Self-healing silk from the sea: role of helical hierarchical structure inPinna nobilisbyssus mechanics

2019

11 pages; International audience; The byssus fibers of Mytilus mussel species have become an important role model in bioinspired materials research due to their impressive properties (e.g. high toughness, self-healing); however, Mytilids represent only a small subset of all byssus-producing bivalves. Recent studies have revealed that byssus from other species possess completely different protein composition and hierarchical structure. In this regard, Pinna nobilis byssus is especially interesting due to its very different morphology, function and its historical use for weaving lightweight golden fabrics, known as sea silk. P. nobilis byssus was recently discovered to be comprised of globula…

Shell matrices of recent rhynchonelliform brachiopods: microstructures and glycosylation studies.

2007

ABSTRACTLike most metazoan biomineralisations, the brachiopod shell is the end product of a biologically controlled calcification process. The main agent of the control is the extracellular matrix, which is secreted by the outer mantle epithelium. This matrix mediates the calcification process by allowing crystal nucleation and elongation in specific orientations and finally, by stopping crystal growth. The proteinaceous moiety of brachiopod shell matrices has been extensively studied. Less known are the post-translational modifications that occur in these matrices, in particular glycosylations. In this comparison of five species of Recent articulated brachiopods, the ratio of soluble to in…

Proteomic analysis of the organic matrix of the abalone Haliotis asinina calcified shell.

2010

Abstract Background The formation of the molluscan shell is regulated to a large extent by a matrix of extracellular macromolecules that are secreted by the shell forming tissue, the mantle. This so called "calcifying matrix" is a complex mixture of proteins and glycoproteins that is assembled and occluded within the mineral phase during the calcification process. While the importance of the calcifying matrix to shell formation has long been appreciated, most of its protein components remain uncharacterised. Results Recent expressed sequence tag (EST) investigations of the mantle tissue from the tropical abalone (Haliotis asinina) provide an opportunity to further characterise the proteins …

Mollusc shellomes: Past, present and future.

2020

13 pages; International audience; In molluscs, the shell fabrication requires a large array of secreted macromolecules including proteins and polysaccharides. Some of them are occluded in the shell during mineralization process and constitute the shell repertoire. The protein moieties, also called shell proteomes or, more simply, 'shellomes', are nowadays analyzed via high-throughput approaches. These latter, applied so far on about thirty genera, have evidenced the huge diversity of shellomes from model to model. They also pinpoint the recurrent presence of functional domains of diverse natures. Shell proteins are not only involved in guiding the mineral deposition, but also in enzymatic a…

Staining SDS-PAGE gels of skeletal matrices after western blot: a way to improve their sharpness.

2015

7 pages; International audience; Denaturing 1D electrophoresis on acrylamide gels - also referred as SDS-PAGE - is a classical technique for fractionating and visualizing the macromolecular constituents of matrices associated to calcified tissues. This technique has been widely used in association with the subsequent silver nitrate staining. But because matrices associated to calcified tissues are very often glycosylated and constituted of numerous polydisperse macromolecules, the obtained pattern is frequently 'smeary' and discrete bands, when present on the gel, are often blurred, thickened or totally masked by the polydisperse macromolecules. In this paper, we present a simple protocol t…

Acidic shell proteins of the Mediterranean fan mussel Pinna nobilis.

2011

43 pages; In molluscs, the shell secretion process is controlled by a set of extracellular macromolecules collectively called the shell matrix. The shell matrix, which is produced by the mantle epithelial cells during mineralization, is predominantly composed of proteins, glycoproteins, acidic polysaccharides, and chitin that precisely regulate the deposition of calcium carbonate outside the mantle cells. In the present paper, we focus on the shell of Pinna nobilis, the giant Mediterranean fan mussel, usually considered as a model for studying molluscan biomineralization processes. P. nobilis exhibits indeed a nacro-prismatic shell, the outer layer of which is constituted of the so-called "…

Acidic Monosaccharides become Incorporated into Calcite Single Crystals*.

2020

Streptococcus suis is an encapsulated bacterium and one of the most important swine pathogens and a zoonotic agent for which no effective vaccine exists. Bacterial capsular polysaccharides (CPSs) are poorly immunogenic, but anti-CPS antibodies are essential to the host defense against encapsulated bacteria. In addition to the previously known serotypes 2 and 14, that are non-immunogenic, we have recently purified and described the CPS structures for serotypes 1, 1/2, 3, 7, 8, and 9. Here, we aimed to elucidate how these new structurally diverse CPSs interact with the immune system to generate anti-CPS antibody responses. CPS-stimulated dendritic cells produced significant levels of C–C moti…

Identification of Two Carbonic Anhydrases in the Mantle of the European Abalone Haliotis tuberculata (Gastropoda, Haliotidae): Phylogenetic Implicati…

2012

Carbonic anhydrases (CAs) represent a diversified family of metalloenzymes that reversibly catalyze the hydration of carbon dioxide. They are involved in a wide range of functions, among which is the formation of CaCO(3) skeletons in metazoans. In the shell-forming mantle tissues of mollusks, the location of the CA catalytic activity is elusive and gives birth to contradicting views. In the present paper, using the European abalone Haliotis tuberculata, a key model gastropod in biomineralization studies, we identified and characterized two CAs (htCA1 and htCA2) that are specific of the shell-forming mantle tissue. We analyzed them in a phylogenetic context. Combining various approaches, inc…

Skeletal organic matrices in molluscs: origin, evolution, diagenesis

2017

8 pages; International audience; The mollusc shell comprises a small amount of organic macromolecules, mostly proteins and polysaccharides, which, all together, constitute the skeletal organic matrix (SOM). In the recent years, the study of the SOM of about two dozens of mollusc species via transcriptomics and/or proteomics has led to the identification of hundreds of shell-associated proteins. This rapidly growing set of data allows several comparisons, shedding light on similarities and differences at the primary structure level and on some peculiar evolutionary mechanisms that may have affected SOM proteins. In addition, it constitutes a prerequisite for investigating the SOM repertoires…

Biochemical characteristics of the shell soluble organic matrix of some recent Rhynchonelliformea (Brachiopoda).

2005

11 pages; International audience; To build up their shell, brachiopods secrete a mixture of proteins and polysaccharides, collectively called the shell organic matrix. This matrix is supposed to mediate the calcification process by allowing the crystal nucleation and elongation and finally, by stopping the crystal growth. The aim of this work is, firstly, to observe at nanoscale the close relationships between the mineral and the organic phases, and secondly, to characterize the acetic acid-soluble matrices extracted from different articulate brachiopods. Recent articulate brachiopod shells were observed according to a standard procedure, which emphasizes the microstructural details. For th…

Metazoan calcium carbonate biomineralizations: macroevolutionary trends – challenges for the coming decade

2014

AbstractCalcium carbonate-based biominerals, also referred as biocalcifications, are the most abundant biogenic mineralized products at the surface of the Earth. In this paper, we summarize general concepts on biocalcifications and we sketch macro-evolutionary trends throughout the history of the Earth, from Archean to Phanerozoic times. Then, we expose five fundamental issues that represent key-challenges in biocalcification researches for the coming decade: the first one concerns the comprehension of the micro- and nano-structure of calcium carbonate biominerals from a mineral viewpoint, while the second one deals with the understanding of the dynamic process of their fabrication. The thi…

Proteomic analysis of the acid-soluble nacre matrix of the bivalve Unio pictorum: detection of novel carbonic anhydrase and putative protease inhibit…

2010

10 pages; International audience; The matrix extracted from mollusc shell nacre is a mixture of proteins and glycoproteins that is thought to play a major role in controlling biomineral synthesis and in increasing its mechanical properties. We investigated the nacreous shell of the freshwater mussel Unio pictorum, to which we applied a proteomics approach adapted to mollusc shell proteins. On one hand, the acid-soluble nacre matrix was fractionated by SDS-PAGE and the five main protein bands (P95, P50, P29, P16, and P12) were digested with trypsin and analyzed by nanoLC-MS/MS followed by de novo sequencing. On the other hand, the acid-soluble nacre matrix was analyzed in a similar manner, w…

Protein-induced, previously unidentified twin form of calcite.

2007

Using single-crystal x-ray diffraction, we found a formerly unknown twin form in calcite crystals grown from solution to which a mollusc shell-derived 17-kDa protein, Caspartin, was added. This intracrystalline protein was extracted from the calcitic prisms of the Pinna nobilis shells. The observed twin form is characterized by the twinning plane of the (108)-type, which is in addition to the known four twin laws of calcite identified during 150 years of investigations. The established twin forms in calcite have twinning planes of the (001)-, (012)-, (104)-, and (018)-types. Our discovery provides additional evidence on the crucial role of biological macromolecules in biomineralization.

Molluscan biomineralization: The proteinaceous shell constituents of Pinna nobilis L.

2005

Abstract The shell of molluscs is a remarkable example of a natural composite biomaterial, synthesized at ambient temperature. Consequently, many consider it as a model for trying to develop at little cost new biomimetic materials of superior mechanical properties. The peculiar resistance of shells to fracture lies in an organic matrix, which is closely associated with the mineral phase. This matrix regulates the crystal growth, by allowing nucleation of the crystals only where appropriate, by favoring crystal elongation in privileged directions, and by inhibiting their growth. This matrix is a mixture of glycoproteins and polysaccharides, the primary structure and function of which are poo…

Author response: 'Palaeoshellomics’ reveals the use of freshwater mother-of-pearl in prehistory

2019

The formation and mineralization of mollusk shell.

2011

27 pages; International audience; In the last years, the field of mollusk biomineralization has known a tremendous mutation. The most recent advances deal with the nanostructure of shell biominerals, and with the identification of several shell matrix proteins: on one hand, the complex hierarchical organization of shell biominerals has been deciphered in few models, like nacre. On the other hand, although proteins represent a minor shell component, they are the major macromolecules that control biocrystal synthesis. Until recently, the paradigm was to consider that this control occurs by two antagonist mechanisms: crystal nucleation and growth inhibition. Emerging models try to translate a …

Calcification in the Shell of the Freshwater Bivalve Unio pictorum.

2005

8 pages; International audience; The scope of the present study is to elucidate the primary structure of the shell proteins of the mollusk Unio pictorum. This freshwater bivalve exhibits an aragonitic shell with nacro-prismatic microstructures. The acetic acid-soluble matrix of the nacreous layer has been extracted, and analyzed on standard polyacrylamide gel under denaturing conditions. Five main proteins can be observed. Their respective apparent molecular weight is 95, 50, 29, 14 and 10 kDa. The three high molecular weight bands are acidic, glycosylated and bind calcium in vitro. These proteins are purified by preparative SDSPAGE. In addition, these fractions are tested to check their ab…

The shell matrix of the pulmonate land snail Helix aspersa maxima.

2012

12 pages; International audience; In mollusks, the shell mineralization process is controlled by an array of proteins, glycoproteins and polysaccharides that collectively constitute the shell matrix. In spite of numerous researches, the shell protein content of a limited number of model species has been investigated. This paper presents biochemical data on the common edible land snail Helix aspersa maxima, a model organism for ecotoxicological purposes, which has however been poorly investigated from a biomineralization viewpoint. The shell matrix of this species was extracted and analyzed biochemically for functional in vitro inhibition assay, for amino acid and monosaccharides composition…

'Shellome': Proteins involved in mollusk shell biomineralization - diversity, functions.

2011

18 pages; International audience

Evolution and biomineralization of pteropod shells

2021

12 pages; International audience; Shelled pteropods, known as sea butterflies, are a group of small gastropods that spend their entire lives swimming and drifting in the open ocean. They build thin shells of aragonite, a metastable polymorph of calcium carbonate. Pteropod shells have been shown to experience dissolution and reduced thickness with a decrease in pH and therefore represent valuable bioindicators to monitor the impacts of ocean acidification. Over the past decades, several studies have highlighted the striking diversity of shell microstructures in pteropods, with exceptional mechanical properties, but their evolution and future in acidified waters remains uncertain. Here, we re…

Caspartin and calprismin, two proteins of the shell calcitic prisms of the Mediterranean fan mussel Pinna nobilis.

2005

We used the combination of preparative electrophoresis and immunological detection to isolate two new proteins from the shell calcitic prisms of Pinna nobilis, the Mediterranean fan mussel. The amino acid composition of these proteins was determined. Both proteins are soluble, intracrystalline, and acidic. The 38-kDa protein is glycosylated; the 17-kDa one is not. Ala, Asx, Thr, and Pro represent the dominant residues of the 38-kDa protein, named calprismin. An N-terminal sequence was obtained from calprismin. This sequence, which comprises a pattern of 4 cysteine residues, is not related to any known protein. The second protein, named caspartin, exhibits an unusual amino acid composition, …

The shell matrix of the freshwater mussel Unio pictorum (Paleoheterodonta, Unionoida). Involvement of acidic polysaccharides from glycoproteins in na…

2007

13 pages; International audience; Among molluscs, the shell biomineralization process is controlled by a set of extracellular macromolecular components secreted by the calcifying mantle. In spite of several studies, these components are mainly known in bivalves from only few members of pteriomorph groups. In the present case, we investigated the biochemical properties of the aragonitic shell of the freshwater bivalve Unio pictorum (Paleoheterodonta, Unionoida). Analysis of the amino acid composition reveals a high amount of glycine, aspartate and alanine in the acid-soluble extract, whereas the acid-insoluble one is rich in alanine and glycine. Monosaccharidic analysis indicates that the in…

Shell repair process in the green ormer Haliotis tuberculata: a histological and microstructural study.

2008

In the present paper, juvenile and adult shells of the green ormer Haliotis tuberculata ('Oreille de Saint-Pierre') were perforated in a zone close to the shell edge and the shell repair process was followed at two levels: (1) by observing the histology of the calcifying mantle in the repair zone and (2) by analyzing with SEM the microstructure of the shell repair zone. Histological data clearly show the presence of calcium carbonate granules into the connective tissues, but not in the epithelial cells. This suggests that calcium carbonate granules are synthesized by sub-epithelial cells and actively transported through the epithelium to the repair zone, via a process which may be similar t…

Proteins as Functional Units of Biocalcification – An Overview

2016

High-throughput approaches such as genomics, transcriptomics and proteomics have led to the discovery of a larger set of biomineralization genes than previously foreseen. These gene lists are often difficult to decode in light of the current models of calcification. Here we overview the proteins available in UniProt (Universal Protein Resource), that were identified directly in metazoan calcium carbonate mineralized structures or known to have direct key-functions in calcification processes. Functional annotation of the protein datasets using Gene Ontology reveals that functions like carbohydrate binding, structural and catalytic activities (e.g. hydrolase) are commonly represented across t…

Evolution of nacre: biochemistry and proteomics of the shell organic matrix of the cephalopod Nautilus macromphalus.

2009

12 pages; International audience; In mollusks, one of the most widely studied shell textures is nacre, the lustrous aragonitic layer that constitutes the internal components of the shells of several bivalves, a few gastropods, and one cephalopod: the nautilus. Nacre contains a minor organic fraction, which displays a wide range of functions in relation to the biomineralization process. Here, we have biochemically characterized the nacre matrix of the cephalopod Nautilus macromphalus. The acid-soluble matrix contains a mixture of polydisperse and discrete proteins and glycoproteins, which interact with the formation of calcite crystals. In addition, a few bind calcium ions. Furthermore, we h…

<p class="HeadingRunIn"><strong>Reinterpretation of the enigmatic Ordovician genus <em>Bolboporites </em>(Echinodermata)</…

2019

Bolboporites is an enigmatic Ordovician cone-shaped fossil, the precise nature and systematic affinities of which have been controversial over almost two centuries. For the first time, a wide range of techniques (CT-scan, SEM, cathodoluminescence, XPL, UV epifluorescence, EBSD, FT-IR and XRF spectrometry) were applied to well-preserved specimens of Bolboporites from Norway and Russia. Our main finding confirms its echinoderm affinities, as shown by its stereomic microstructure and by the first definitive evidence of its monocrystalline nature. Each cone consists in a single, microporous calcitic crystal with a narrow longitudinal internal canal. These results are combined with all previous …

Merging models of biomineralisation with concepts of nonclassical crystallisation: is a liquid amorphous precursor involved in the formation of the p…

2012

16 pages; International audience; The calcitic prisms of Pinna nobilis (Pinnidae, Linnaeus 1758) are shown to be perfect examples of a mesocrystalline material. Based on their ultrastructure and on the occurrence of an amorphous transient precursor during the early stages of prism formation, we provide evidence for the pathway of mesocrystallisation proposed by Seto et al. (2012), which proceeds not by self-organized oriented attachment of crystalline nano-bricks but by aggregation of initially amorphous nanogranules which later transform by epitaxial nucleation to a threedimensional array of well aligned nanocrystals. We further fathom the role of a liquid amorphous calcium carbonate in bi…

Experimental modeling of bacterially-induced Ca carbonate precipitation: new insights on possible mechanisms.

2015

The contribution of microorganisms, particularly bacteria, in carbonate mineral formation, the main natural processes controlling CO2 level in the atmosphere, has played an important role since the Archean Eon. In this study we review our recent experimental work on CaCO3 precipitation induced by two anoxygenic phototrophic bacteria (APB), Rhodovulum steppense A-20sT and Rhodovulum sp. S-17-65, and by cyanobacteria Gloeocapsa sp. f-6gl. These bacteria are representatives of two important groups of photosynthetic organisms present at the Earth surface both in the past and at the present times. The mechanisms of organomineralization deriving from APB and cyanobacteria activities are drastical…

Synthesis of calcium carbonate biological materials: how many proteins are needed?

2013

In Nature, calcium carbonate biomineralizations are the most abundant mineralized structures of biological origin. Because many exhibit remarkable characteristics, several attempts have been made to use them as substitution materials for bone reconstruction or as models for generating biomimetic composites that exhibit tailored properties. CaCO3biomineralizations contain small amounts of amalgamate of proteins and polysaccharides that are secreted during the calcification process. They contribute to control the morphology of the crystallites and to spatially organize them in well-defined microstructures. These macromolecules, collectively defined as the skeletal matrix, have been the focus …

Characterization of the Teeth Skeletal Matrix from <i>Arbacia lixula</i>

2016

The teeth of sea urchins are highly complex composite structures, composed predominantly of high magnesium calcite, and of a minor heterogeneous assemblage of organic macromolecules that are occluded within the mineral. The organic matrix fulfils important functions in mineralization, in addition to giving the mineral phase peculiar mechanical properties, different from that of purely inorganic calcite. Nevertheless, the composition and function of individual components of the organic matrix still remains largely unknown. Up to now, the detailed protein repertoire of teeth from a single sea urchin species (Strongylocentrotus purpuratus, order Camarodonta) was investigated. In this study, we…

Novel molluskan biomineralization proteins retrieved from proteomics: a case study with upsalin.

2012

12 pages; International audience; The formation of the molluskan shell is regulated by an array of extracellular proteins secreted by the calcifying epithelial cells of the mantle. These proteins remain occluded within the recently formed biominerals. To date, many shell proteins have been retrieved, but only a few of them, such as nacreins, have clearly identified functions. In this particular case, by combining molecular biology and biochemical approaches, we performed the molecular characterization of a novel protein that we named Upsalin, associated with the nacreous shell of the freshwater mussel Unio pictorum. The full sequence of the upsalin transcript was obtained by RT-PCR and 5'/3…

Unveiling the evolution of bivalve nacre proteins by shell proteomics of Unionoidae.

2015

The formation of the molluscan shell nacre is regulated to a large extent by a matrix of extracellular macromolecules that are secreted by the shell forming tissue, the mantle. This so called “calcifying matrix” is a complex mixture of proteins and glycoproteins that is assembled and occluded within the mineral phase during the calcification process. While the importance of the calcifying matrix to shell formation has long been appreciated, the molecular basis that dictates nacre formation remains largely uncharacterized.Recent expressed sequence tag (EST) investigations of the freshwater mussels (Elliptio complanata and Villosa leinosa) provide an opportunity to further characterize the pr…

Characterization of crustacyanin-A2 subunit as a component of the organic matrix of gastroliths from the crayfish Cherax quadricarinatus.

2009

AbstractLike the lobsters, some terrestrial crabs and other crayfishes, the Australian red claw crayfish, Cherax quadricarinatus, elaborates in its stomach wall calcium storage structures called gastroliths. For understanding the cyclic elaboration and stabilization of these amorphous calcified structures, we studied the organic matrix (OM) of these paired biomineralizations. After decalcification with acetic acid, we analysed the proteinaceous components of an acetic acid-insoluble fraction by two-dimensional electrophoresis. Nine spots were digested by trpsin and the tryptic peptides were sequenced by nanoLC-nanoESI-MS/MS mass spectrometry. About 100 peptidic sequences were compared to se…

Shell Extracts from the Marine Bivalve Pecten maximus Regulate the Synthesis of Extracellular Matrix in Primary Cultured Human Skin Fibroblasts

2014

International audience; Mollusc shells are composed of more than 95% calcium carbonate and less than 5% of an organic matrix consisting mostly of proteins, glycoproteins and polysaccharides. Previous studies have elucidated the biological activities of the shell matrices from bivalve molluscs on skin, especially on the expression of the extracellular matrix components of fibroblasts. In this work, we have investigated the potential biological activities of shell matrix components extracted from the shell of the scallop Pecten maximus on human fibroblasts in primary culture. Firstly, we demonstrated that shell matrix components had different effects on general cellular activities. Secondly, …

Shell proteome of rhynchonelliform brachiopods.

2015

7 pages; International audience; Brachiopods are a phylum of marine invertebrates that have an external bivalved shell to protect their living tissues. With few exceptions, this biomineralized structure is composed of calcite, mixed together with a minor organic fraction, comprising secreted proteins that become occluded in the shell structure, once formed. This organic matrix is thought to display several functions, in particular, to control mineral deposition and to regulate crystallite shapes. Thus, identifying the primary structure of matrix proteins is a prerequisite for generating bioinspired materials with tailored properties. In this study, we employed a proteomic approach to identi…

Archéologie - Coquillages locaux.

2019

International audience

The role of enzymes in biomineralization processes.

2008

56 pages; International audience

Localization of biomineral proteins by Atomic Force Microscopy (AFM) with functionalized tips

2021

Accretion in strong field gravity with eXTP

2019

In this paper we describe the potential of the enhanced X-ray Timing and Polarimetry (eXTP) mission for studies related to accretion flows in the strong field gravity regime around both stellar-mass and supermassive black-holes. eXTP has the unique capability of using advanced 'spectral-timing-polarimetry' techniques to analyze the rapid variations with three orthogonal diagnostics of the flow and its geometry, yielding unprecedented insight into the inner accreting regions, the effects of strong field gravity on the material within them and the powerful outflows which are driven by the accretion process.

The evolution of metazoan α-carbonic anhydrases and their roles in calcium carbonate biomineralization

2014

The carbonic anhydrase (CA; EC 4.2.1.1) superfamily is a class of ubiquitous metallo-enzymes that catalyse the reversible hydration of carbon dioxide. The ?-CA family, present in all metazoan clades, is a key enzyme involved in a wide range of physiological functions including pH regulation, respiration, photosynthesis, and biocalcification. This paper reviews the evolution of the ?-CA family, with an emphasis on metazoan ?-CA members involved in biocalcification. Phylogenetic analyses reveal a complex evolutionary history of ?-CAs, and suggest ?-CA was independently co-opted into a variety of skeleton forming roles (e.g. as a provider of HCO3? ions, a structural protein, a nucleation activ…

Preface

2015

Reevaluation of the enigmatic Ordovician genus Bolboporites.

2018

International audience

Supplementary figures from Deep conservation of bivalve nacre proteins highlighted by shell matrix proteomics of the Unionoida Elliptio complanata an…

2017

figures S1-S3

Il y a nacre et nacre.

2016

1 page - commentaire; National audience

La "Paléoshellomique" dans les starting-blocks

2021

Supplementary table from Deep conservation of bivalve nacre proteins highlighted by shell matrix proteomics of the Unionoida Elliptio complanata and …

2017

Table S1