0000000000122142
AUTHOR
Gilles Luquet
showing 28 related works from this author
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…
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…
Proteoglycan occurrence in gastrolith of the crayfish Cherax quadricarinatus (Malacostraca: Decapoda).
2012
14 pages; International audience; Biomineralized structures are hybrid composites formed and stabilized by the close interaction of the organic and the inorganic phases. Crayfish are good models for studying biomineralization because they develop, in a molting-mineralization cycle, semi-spherical mineralized structures referred to as gastroliths. The organic matrix of these structures consists of proteins, polysaccharides, and lipids. Chitin is the main polysaccharide and is concentrically arranged as fibrous chitin-protein lamellar structures. Although several proteins and low-molecular weight phosphorylated components have been reported to be involved in gastrolith mineralization, the occ…
Biomineralizations: insights and prospects from crustaceans.
2011
19 pages; International audience; For growing, crustaceans have to molt cyclically because of the presence of a rigid exoskeleton. Most of the crustaceans harden their cuticle not only by sclerotization, like all the arthropods, but also by calcification. All the physiology of crustaceans, including the calcification process, is then linked to molting cycles. This means for these animals to find regularly a source of calcium ions quickly available just after ecdysis. The sources of calcium used are diverse, ranging from the environment where the animals live to endogenous calcium deposits cyclically elaborated by some of them. As a result, crustaceans are submitted to an important and energ…
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…
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 …
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,…
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…
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…
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…
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 …
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…
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…
Stable amorphous calcium carbonate is the main component of the calcium storage structures of the crustacean Orchestia cavimana.
2002
Amorphous calcium carbonate (ACC) is the least stable form of the six known phases of calcium carbonate. It is, however, produced and stabilized by a variety of organisms. In this study we examined calcium storage structures from the terrestrial crustacean Orchestia cavimana, in order to better understand their formation mode and function. By using X-ray diffraction, infrared and Raman spectroscopy, thermal analysis and elemental analysis, we determined that the mineral comprising these storage structures is amorphous calcium carbonate with small amounts of amorphous calcium phosphate (5%). We suggest that the use of amorphous calcium carbonate might be advantageous for these storage struct…
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…
Orchestin, a calcium-binding phosphoprotein, is a matrix component of two successive transitory calcified biomineralizations cyclically elaborated by…
2003
Orchestia cavimana is a crustacean that cyclically replaces its calcified cuticle during molting cycles in order to grow. Its terrestrial way of life requires storage of calcium during each premolt period, as calcareous concretions, in tubular diverticula of the midgut. During the postmolt period the stored calcium is reabsorbed and is translocated through the storage organ epithelium as calcified small spherules. In a previous study, we sequenced and characterized a remarkable component of the organic matrix of the premolt storage structures, Orchestin, which is a calcium-binding phosphoprotein. In this paper, we analyzed the spatiotemporal expression of the orchestin gene by Northern blot…
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…
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…
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…
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…
Comparative ultrastructure and carbohydrate composition of gastroliths from Astacidae, Cambaridae and Parastacidae freshwater crayfish (Crustacea, De…
2012
21 pages; International audience; Crustaceans have to cyclically replace their rigid exoskeleton in order to grow. Most of them harden this skeleton by a calcification process. Some decapods (land crabs, lobsters and crayfish) elaborate calcium storage structures as a reservoir of calcium ions in their stomach wall, as so-called gastroliths. For a better understanding of the cyclic elaboration of these calcium deposits, we studied the ultrastructure of gastroliths from freshwater crayfish by using a combination of microscopic and physical techniques. Because sugars are also molecules putatively involved in the elaboration process of these biomineralizations, we also determined their carbohy…
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…
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…
Influence of female moulting status on pairing decisions and size‐assortative mating in amphipods
2012
8 pages; International audience; Precopulatory mate guarding is a common strategy, which has evolved in species where the female receptivity (and thus egg fertilization) is predictable, but also limited to a short period. Although males are larger than females in many amphipods, the largest males pair with the largest females, leading to a positive sizeassortative pairing. Size-assortative pairing has received much attention but how moulting physiology could affect pairing decisions has rarely been studied. Here, we tested the hypothesis that the size-assortative pairing in the freshwater amphipod Gammarus pulex is closely related to the female moult cycle. We characterized moulting status …