Proteomic Analysis of the Acid-Soluble Nacre Matrix of the Bivalve Unio pictorum: Detection of Novel Carbonic Anhydrase and Putative Protease Inhibitor Proteins
IBCP;
Université de Bourgogne;
6 Boulevard Gabriel;
UMR 5561 CNRS Biogéosciences;
21000 Dijon (France);
Institut de Biologie et Chimie des Protéines;
IFR 128 BioSciences Gerland-Lyon Sud;
UMR 5086 CNRS;
69367 Lyon (France);
Université de Lyon 1;
7 passage du Vercors;
关键词：
biomineralization;
proteomics;
de novo sequencing;
genomics;
glycoproteins;
期刊名称：
Chembiochem: A European journal of chemical biology
i s s n：
1439-4227
年卷期：
2010 年
11 卷
15 期
页 码：
2138-2147
页 码：
摘 要：
Abstract 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, without any preliminary fractionation. In total, we obtained about 140 peptides, of between 9 and 21 residues, as well as several shorter peptides. Interestingly, it appears that the different protein bands share several identical peptides; this has implications for the underlying genetic machinery that synthesizes nacre proteins. Homology searches against sequences in the Swiss-Prot protein database and the 800?000 mollusc expressed sequence tag database were performed, but surprisingly, only a few obvious homologies were established. Among the peptides that match with known sequences, some from P50 and P16/P12 proteins align with carbonic anhydrase (CA) and with the protease inhibitor, respectively. The evolutionary implications of our findings are discussed.
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