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Novel Proteins from the Calcifying Shell Matrix of the Pacific Oyster Crassostrea gigas

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Abstract

The shell of the Pacific oyster Crassostrea gigas is composed of more than 99% CaCO3 and of around 0.5% of occluded organic matrix. According to classical views, this matrix is supposed to regulate the shell mineral deposition. In this study, we developed one of the first proteomic approaches applied to mollusk shell in order to characterise the calcifying matrix proteins. The insoluble organic matrix, purified after demineralisation of the shell powder, was digested with trypsin enzyme, and separated on nano-LC, prior to nanospray quadrupole/time-of-flight analysis. MS/MS spectra were searched against the above 220,000 EST sequences available in the public database for Crassostrea. Using this approach, we were able to identify partial or full-length sequence transcripts that encode eight novel shell matrix proteins.

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Abbreviations

AA:

Amino acid

ASM:

Acid-soluble matrix

AIM:

Acid-insoluble matrix

EGF:

Epidermal growth factor

IMSP:

Insoluble matrix shell protein

SEM:

Scanning electron microscopy

EST:

Expressed sequence tag

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Acknowledgements

The work of B. Marie and F. Marin is financially supported by an ANR (ACCRO-EARTH, ref. BLAN06-2_159971, coordinator Gilles Ramstein, LSCE) during the period 2007–2011. A complementary financial support was provided by INSU (Action INTERVIE 2010) and by COST action TD0903 (Davorin Medakovic 2009–2013). The “Conseil Régional de Bourgogne” (Dijon, France) provided additional supports for the acquisition of new equipment in the Biogeosciences research unit (tabletop SEM microscope, Hitachi TM1000). B. Marie thanks Jérome Thomas for handling shell picture. The present protein sequences of IMSP-1, IMSP-2, IMSP-3, IMSP4, IMSP-5 and IMSP-6 appear in the UniProtKB under accession no. P86784-P86789 and were also named Gigasin-1, Gigasin-2, Gigasin-3, Gigasin-4, Gigasin-5 and Gigasin-6, respectively.

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Authors and Affiliations

  1. UMR 5561 CNRS Biogéosciences, Université de Bourgogne, 6 Bd. Gabriel, Dijon, 21000, France

    Benjamin Marie, Nathalie Guichard & Frédéric Marin

  2. Université de Bourgogne, Dijon, France

    Benjamin Marie, Nathalie Guichard & Frédéric Marin

  3. IFR 128 BioSciences; UMR 5086 CNRS; IBCP, Université de Lyon 1, Lyon, France

    Isabelle Zanella-Cléon & Michel Becchi

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  1. Benjamin Marie
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  2. Isabelle Zanella-Cléon
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  3. Nathalie Guichard
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  4. Michel Becchi
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  5. Frédéric Marin
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Correspondence to Benjamin Marie or Frédéric Marin.

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Marie, B., Zanella-Cléon, I., Guichard, N. et al. Novel Proteins from the Calcifying Shell Matrix of the Pacific Oyster Crassostrea gigas . Mar Biotechnol 13, 1159–1168 (2011). https://doi.org/10.1007/s10126-011-9379-2

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  • DOI: https://doi.org/10.1007/s10126-011-9379-2

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