Get access

Coupling Proteomics and Transcriptomics for the Identification of Novel and Variant Forms of Mollusk Shell Proteins: A Study with P. margaritifera

Authors

  • Dr. Sophie Berland,

    Corresponding author
    1. UMR BOREA (Biologie des Organismes et Ecosystèmes Aquatiques), MNHN/CNRS 7208/IRD 207, CP 26, 43 rue Cuvier, 75231 Paris Cedex 05 (France), Fax: (+33) 1-40-79-37-71
    • UMR BOREA (Biologie des Organismes et Ecosystèmes Aquatiques), MNHN/CNRS 7208/IRD 207, CP 26, 43 rue Cuvier, 75231 Paris Cedex 05 (France), Fax: (+33) 1-40-79-37-71
    Search for more papers by this author
  • Dr. Arul Marie,

    1. Département RDDM, FRE 3206 CNRS, Plateforme de Spectrométrie de Masse et de Protéomique, Muséum National d'Histoire Naturelle, 57 rue Cuvier, 75005 Paris (France)
    Search for more papers by this author
  • Dr. Denis Duplat,

    1. UMR BOREA (Biologie des Organismes et Ecosystèmes Aquatiques), MNHN/CNRS 7208/IRD 207, CP 26, 43 rue Cuvier, 75231 Paris Cedex 05 (France), Fax: (+33) 1-40-79-37-71
    Search for more papers by this author
  • Dr. Christian Milet,

    1. UMR BOREA (Biologie des Organismes et Ecosystèmes Aquatiques), MNHN/CNRS 7208/IRD 207, CP 26, 43 rue Cuvier, 75231 Paris Cedex 05 (France), Fax: (+33) 1-40-79-37-71
    Search for more papers by this author
  • Dr. Jean Yves Sire,

    1. UMR 7138, Equipe “Evolution et Développement du Squelette”, Université Pierre & Marie Curie, 7 quai St. Bernard, 75005 Paris (France)
    Search for more papers by this author
  • Dr. Laurent Bédouet

    1. UMR BOREA (Biologie des Organismes et Ecosystèmes Aquatiques), MNHN/CNRS 7208/IRD 207, CP 26, 43 rue Cuvier, 75231 Paris Cedex 05 (France), Fax: (+33) 1-40-79-37-71
    Search for more papers by this author

Abstract

Shell matrix proteins from Pinctada margaritifera were characterized by combining proteomics analysis of shell organic extracts and transcript sequences, both obtained from the shell-forming cell by using the suppression subtractive hybridization method (SSH) and from an expressed sequence tag (EST) database available from Pinctada maxima mantle tissue. Some of the identified proteins were homologues to proteins reported in other mollusk shells, namely lysine-rich matrix proteins (KRMPs), shematrins and molluscan prismatic and nacreous layer 88 kDa (MPN88). Sequence comparison within and among Pinctada species pointed to intra- and interspecies variations relevant to polymorphism and to evolutionary distance, respectively. In addition, a novel shell matrix protein, linkine was identified. BLAST analysis of the peptide sequences obtained from the shell of P. margaritifera against the EST database revealed the presence of additional proteins: two proteins similar to the Pif97 protein that was identified in the shell of P. fucata, a chitinase-like protein previously identified in Crassostrea gigas, two chitin-binding proteins, and two incomplete sequences of proteins unknown so far in mollusk shells. Combining proteomics and transcriptomics analysis we demonstrate that all these proteins, including linkine, are addressed to the shell. Retrieval of motif-forming sequences, such as chitin-binding, with functional annotation from several peptides nested in the shell could indicate protein involvement in shell patterning.

Ancillary