Search results for "hemocyanin"
showing 10 items of 148 documents
Molecular Structure of the Arthropod Hemocyanins
1992
Hemocyanin is an extracellular, blue protein that occurs in high concentrations in the blood of many arthropods, including spiders, scorpions, horseshoe crabs, crustaceans, and at least two centipedes. Serving as an ### oxygen carrier, it is functionally equivalent to hemoglobin, but performs reversible oxygen binding between two copper ions. Hemocyanin is composed of a number of subunits that assemble in an extremely large macro-molecular entity. These particles, which are similar in size to viruses or ribosomes, exhibit a complex allosteric behavior during oxygen binding. There is growing evidence that this functional plasticity has evolved upon, and answers to, ecophysiological constrain…
Evolutionary history and diversity of arthropod hemocyanins
2004
Hemocyanins are copper-containing, multi-subunit proteins that transport oxygen in the hemolymph of many molluscs and arthropods [Markl and Decher, Adv. Comp. Environ. Physiol. 13 (1992) 325; van Holde et al., J. Biol. Chem. 276 (2001) 15563]. Arthropod hemocyanins originated more than 550 million years ago from oxygen-consuming phenoloxidases. Hemocyanins are present in various Onychophora, Chelicerata, Myriapoda, Crustacea, and Hexapoda, but subunit evolution differs striking in these arthropod subphyla. Hemocyanins also gave rise to non-respiratory proteins (crustacean pseudo-hemocyanins, insect hexamerins, and hexamerin receptors), which most likely have storage functions.
Immunoelectron Microscopy of Hemocyanin from the Keyhole Limpet (Megathura crenulata): A Parallel Subunit Model
1993
Abstract Immunoelectron microscopy has been performed using negatively stained immune complexes of keyhole limpet hemocyanin (KLH) subunit 2 di- and multidecamers with domain-specific monoclonal antibodies. One antibody (KLH2 a macr 1) links the hemocyanin molecules in a side-to-side pattern, whereas the other antibody (KLH2 fg macr 1) links the molecules end-to-end. From existing knowledge of the domain sequence of KLH subunit 2, these data provide support for a parallel arrangement of subunits within each decamer. Ten N-terminal a macr: domains are then present at the noncollar region of each decamer with 10 C-terminal g macr domains at the collar region. The immunonegative staining data …
Introduction: Protein Oligomerization and the Formation of Macromolecular Assemblies
2019
The ability of biomolecules to link together to form higher order assemblies underlies much of cellular structure and function. Here we emphasise protein oligomerisation and discuss some of the principles of molecular interaction, from early considerations through to the present day. A few protein examples are presented, selected from our research interests, to highlight assembly features, ranging from the hemoglobins, the hemocyanins to the peroxiredoxins, collagen, the encapsulins and ferritins.
Keyhole Limpet Hemocyanin (KLH): Slow In Vitro Reassociation of KLH1 and KLH2 from Immucothel®
1998
Abstract Following our in vitro reassociation of keyhole limpet hemocyanin subunits in the presence of high concentrations (100 mM each) of calcium and magnesium chloride (Harris et al., 1997a, Micron 28, 31–41; 1997b, Micron 28, 43–56), we have now extended our investigations by using a buffer system containing a lower concentration of the two divalent cations (10 mM each). Reassociation of mixed KLH subunits present in the commercially available product Immucothel® was performed using a standardized buffer solution containing 50 mM Tris–HCl, 150 mM NaCl, 10 mM CaCl2 and 10 mM MgCl2 (pH 7.4) over a minimum period of one week, at 4°C. This solution was selected as being close to our KLH sta…
On the stability of the 24-meric hemocyanin from Eurypelma californicum.
1998
The stability of the 24-meric hemocyanin from Eurypelma californicum towards various denaturants (GdnHCl, urea, urea derivatives and salts of the Hofmeister series) indicates that the quaternary structure is stabilized by hydrophilic and polar forces. Thus, the interaction between the seven different subunit types of this cheliceratan hemocyanin is comparable with that of the closely related crustacean hemocyanins. In contrast, no significant influence of divalent ions such as Ca2+ and Mg2+ on the stability is observed at pH 8.0 and pH 8.5 but not at pH 7.0. Studies, both in the presence of urea and GdnHCl indicate that the denaturation process consists of a dissociation of the oligomeric s…
Controlled cleavage of KLH1 and KLH2 by the V8 protease from Staphylococcus aureus reassociation, electrophoretic and transmission electron microscop…
1999
The reassociation behaviour of protease V8-cleaved peptides from KLH1 and KLH2, the two hemocyanin isoforms from the giant keyhole limpet Megathura crenulata, has been studied by transmission electron microscopy of negatively stained specimens and SDS/PAGE. Reassociation of the complete mixture of protease cleavage products and of combinations of peptide fragments purified by HPLC was performed in the presence of 100 mm CaCl2 and 100 mm MgCl2 at pH 7.4, over a period of 1 to 4 weeks. The V8 protease splits KLH1 into peptide fragments containing the functional units abc, def, defg, defgh, g and h. This mixture of peptide fragments reassociated to form helical tubular polymers, with a diamete…
Copper Proteins with Dinuclear Active SitesBased in part on the article Copper Proteins with Dinuclear Active Sites by Konrad Lerch which appeared in…
2006
Copper proteins with dinuclear active sites comprise proteins with different structures and functions. The phenoloxidase, tyrosinase, and catecholoxidase are responsible for browning by starting the synthesis of melanin. These enzymes are involved in the primary immune response in invertebrates, plants, fungi as well as in the sclerotization of arthropods. The respiratory proteins hemocyanins are responsible for oxygen transport in some arthropods and molluscs. However, they can be converted to enzymes exhibiting phenoloxidase activity. Based on X-ray structures of hemocyanins and a catecholoxidase, large parts of folding motifs are very similar although the sequence identities are far belo…
Two-photon excitation microscopy of tryptophan-containing proteins.
2002
We have examined the feasibility of observing single protein molecules by means of their intrinsic tryptophan emission after two-photon excitation. A respiratory protein from spiders, the 24-meric hemocyanin, containing 148 tryptophans, was studied in its native state under almost in vivo conditions. In this specific case, the intensity of the tryptophan emission signals the oxygen load, allowing one to investigate molecular cooperativity. As a system with even higher tryptophan content, we also investigated latex spheres covered with the protein avidin, resulting in 340 tryptophans per sphere. The ratio of the fluorescence quantum efficiency to the bleaching efficiency was found to vary b…
A potential role for water in the modulation of oxygen-binding by tarantula hemocyanin
2003
Hemocyanin from the tarantula Eurypelma californicum is a large respiratory protein with an exceptional high cooperativity. In contrast to hemocyanins from other species, no physiological allosteric effectors other than protons have been identified so far for this 24-meric oligomer. Here we report for the first time the mediating effects of water activity on the oxygen binding properties of a hemocyanin. Oxygen binding curves were measured in presence of several concentrations of glycine and sucrose since both substances reduce water activity. A pronounced shift of the p(50) was observed in both cases but in different directions: adding sucrose shifts the p(50) towards lower values whereas …