The shell matrix of the freshwater mussel Unio pictorum (Paleoheterodonta, Unionoida). Involvement of acidic polysaccharides from glycoproteins in nacre mineralization.
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 insoluble matrix comprises a high amount of glucosamine. Furthermore, a high ratio of the carbohydrates of the soluble matrix is sulfated. Electrophoretic analysis of the acid-soluble matrix revealed discrete bands. Stains-All, Alcian Blue, periodic acid/Schiff and autoradiography with (45)Ca after electrophoretic separation revealed three major polyanionic calcium-binding glycoproteins, which exhibit an apparent molecular mass of 95, 50 and 29 kDa, respectively. Two-dimensional gel electrophoresis shows that these bands, provisionally named P95, P50 and P29, are composed of numerous isoforms, the majority of which have acidic isoelectric points. Chemical deglycosylation of the matrix with trifluoromethanesulfonic acid induces a drastic shift of both the apparent molecular mass and the isoelectric point of these matrix components. This treatment induces also a modification of the shape of CaCO(3) crystals grown in vitro and a loss of the calcium-binding ability of two of the main matrix proteins (P95 and P50). Our findings strongly suggest that post-translational modifications display important functions in mollusc shell calcification.
Marie
Benjamin
B
UMR CNRS 5561, Biogéosciences, Université de Bourgogne, 6 boulevard Gabriel, F-21000 Dijon, France. benjamin.marie@u-bourgogne.fr
Luquet
Gilles
G
Pais De Barros
Jean-Paul
JP
Guichard
Nathalie
N
Morel
Sylvain
S
Alcaraz
Gérard
G
Bollache
Loïc
L
Marin
Frédéric
F
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D049872:Bivalvia / Q000737:chemistry*; D002113:Calcification, Physiologic; D002241:Carbohydrates / Q000032:analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000737:chemistry* / Q000502:physiology; D005618:Fresh Water; D006023:Glycoproteins / Q000502:physiology*; D011134:Polysaccharides / Q000502:physiology*; D017550:Spectroscopy, Fourier Transform Infrared
D000596:Amino Acids; D002241:Carbohydrates; D006023:Glycoproteins; D011134:Polysaccharides