Abstract
Nacre of the Pinctada pearl oyster shells is composed of 98% CaCO3 and 2% organic matrix. The relationship between the organic matrix and the mechanism of nacre formation currently constitutes the main focus regarding the biomineralization process. In this study, we isolated a new nacre matrix protein in P. margaritifera and P. maxima, we called Pmarg- and Pmax-MRNP34 (methionine-rich nacre protein). MRNP34 is a secreted hydrophobic protein, which is remarkably rich in methionine, and which is specifically localised in mineralizing the epithelium cells of the mantle and in the nacre matrix. The structure of this protein is drastically different from those of the other nacre proteins already described. This unusual methionine-rich protein is a new member in the growing list of low complexity domain containing proteins that are associated with biocalcifications. These observations offer new insights to the molecular mechanisms of biomineralization.
Similar content being viewed by others
Abbreviations
- AA:
-
Amino-acid
- AIM:
-
Acid-insoluble matrix
- ASM:
-
Acid-soluble matrix
- EST:
-
Expressed sequence tag
- ISH:
-
In situ hybridization
- MRNP:
-
Methionine-rich nacre protein
- MS:
-
Mass spectrometry
- RLCD:
-
Repeated low complexity domain
References
Addadi L, Joester D, Nudelman F, Weiner S (2006) Mollusk shell formation: a source of new concepts for understanding biomineralization processes. Chem Eur J 12:980–987
Cochennec-Laureau N, Montagnani C, Saulnier D, Fougerouse A, Levy P, Lo C (2010) A histological examination of grafting success in pearl oyster Pinctada margaritifera in French Polynesia. Aquatech Living Resour 23:131–140
Cunha R, Blanc F, Bonhomme F, Arnaud-Haond S (2011) Evolutionary pattern in pearl oysters of the genus Pinctada (Bivalvia: Pteriidae). Mar Biotechnol 13:181–192
Falini G, Albeck S, Weiner S, Addadi L (1996) Control of aragonite or calcite polymorphism by mollusk shell macromolecules. Science 271:67–69
Fleury E, Huvet A, Lelong C, de Lorgeril J, Boulo V, Gueguen Y, Bachère E, Tanguy A, Moraga D, Fabioux C, Lindeque P, Shaw J, Reinhardt R, Prunet P, Davey G, Lapègue S, Sauvage C, Corporeau C, Moal J, Gavory F, Wincker P, Moreews F, Klopp C, Mathieu M, Boudry P, Favrel P (2009) Generation and analysis of a 29, 745 unique expressed sequence tags from the Pacific oyster (Crassostrea gigas) assembled into a publicly accessible database: the GigasDatabase. BMC genomics 10:341
Jackson DJ, McDougall C, Woodcroft B, Moase P, Rose RA, Kube M, Reinhardt R, Rokhsar DS, Montagnani C, Joubert C, Piquemal D, Degnan BM (2010) Parallel evolution of nacre building gene sets in molluscs. Mol Biol Evol 27:591–608
Joubert C, Piquemal D, Marie B, Manchon L, Pierrat F, Zanella-Cléon I, Cochennec-Laureau N, Gueguen Y, Montagnani C (2010) Transcriptome and proteome analysis of Pinctada margaritifera calcifying mantle and shell: focus on biomineralization. BMC Genomics 11:613
Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132
Lao Y, Zhang X, Zhou J, Su W, Chen R, Wang Y, Zhou W, Xu ZF (2007) Characterization and in vitro mineralization function of a soluble protein complex P60 from the nacre of Pinctada fucata. Comp Biochem Physiol B 148:201–208
Larsen JB, Frisher ME, Rasmussen J, Hansqen BW (2005) Single-step nested multiplex PCR to differentiate between various bivalve larvae. Mar Biol 146:1119–1129
Lowenstam HA, Weiner S (1989) On biomineralization. Oxford University Press, New York
Mann S (2001) Biomineralization: principles and concepts in bioinorganic materials chemistry. Oxford University Press, New York
Marie B, Luquet G, Pais de Barros J-P, Guichard N, Morel S, Alcaraz G, Bollache L, Luquet G, Marin F (2007) The shell matrix of the freshwater mussel Unio pictorum (paleoheterodonta, unionoida): involvement of acidic polysaccharides from glycoproteins in nacre mineralization. FEBS J 274:2933–2945
Marie B, Luquet G, Bédouet L, Milet C, Guichard N, Médakovic D, Marin F (2008) Nacre calcification in the freshwater mussel Unio pictorum: carbonic anhydrase activity and purification of a 95 kDa calcium-binding glycoprotein. Chem Biochem 9:2515–2523
Marie B, Marie A, Jackson DJ, Dubost L, Degnan BM, Milet C, Marin F (2010a) Proteomic analysis of the organic matrix of the abalone Haliotis asinina calcified shell. Proteome Sci 8:54
Marie B, Zanella-Cléon I, Le Roy N, Becchi M, Luquet G, Marin F (2010b) Proteomic analysis of the acid-soluble nacre matrix of the bivalve Unio pictorum: detection of novel carbonic anhydrase and putative protease inhibitor proteins. Chem Biochem 11:2138–2147
Marin F, Muyzer G, Dauphin Y (1994) Caractérisations électrophorétique et immunologique des matrices organiques solubles des tests de deux bivalves ptériomorphes actuels, Pinna nobilis et Pinctada margaritifera. C R Acad Sci Paris 318:1653–1659
Marin F, Corstjens B, Gaulejac E, De Vring-de Jong E, Westbroek P (2000) Mucins and molluscan calcification - Molecular characterization of mucoperlin, a novel mucin-like protein from the nacreous shell layer of the fan mussel Pinna nobilis (Bivalvia, Pteriomorphia). J Biol Chem 275:20667–20675
Marin F, Pereira L, Westbroek P (2001) Large-scale fractionation of molluscan shell matrix. Protein Expr Purif 23:175–179
Marin F, Luquet G, Marie B, Médakovic D (2008) Molluscan shell proteins: primary structure, origin and evolution. Curr Top Dev Biol 80:209–276
Miyamoto H, Miyashita T, Okushima M, Nakano S, Morita T, Matsushiro (1996) A carbonic anhydrase from the nacreous layer in oyster pearls. Proc Natl Acad Sci 93:9657–9660
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45
Pokroy B, Fitch AN, Marin F, Kapon N, Adir N, Zolotoyabko E (2006) Anisotropic lattice distorsion in biogenic calcite induced by intra-crystalline organic molecules. J Struct Biol 155:96–103
Shen X, Belcher AM, Hansma PK, Stucky GD, Morse DE (1997) Molecular cloning and characterization of lustrin-A: a matrix protein from shell and pearl nacre of Haliotis rufescens. J Biol Chem 272:32472–32481
Siivari K, Zhang M, Palmer AG, Vogel HJ (1995) NMR studies of the methionine methyl groups in calmodulin. FEBS Lett 366:104–108
Sudo S, Fujikawa T, Nagakura T, Ohkubo T, Sakaguchi K, Tanaka M, Nakashima K, Takahashi T (1997) Structure of mollusc shell framework proteins. Nature 387:563–564
Suzuki M, Saruwatari K, Kogure T, Yamamoto Y, Nishimura T, Nishimura T, Kato T, Nagasawa H (2009) An acidic matrix protein, Pif, is a key macromolecules for nacre formation. Science 325:1388–1390
Wheeler AP, George JW, Evans CA (1981) Control of CaCO3 nucleation and crystal growth by soluble matrix of oyster shell. Science 212:1397–1398
Acknowledgments
This study is part of a collaborative project (GDR ADEQUA) supported by the “Service de la perliculture” of French Polynesia. It is also supported by Ifremer, the University of French Polynesia, and the University of Burgundy. Authors are grateful to Nathalie Cochennec-Laureau, Marcel Le Pennec, Péva Levy, Cédrik Lo and Anne-Sandrine Talfer for helpful discussions and assistance. The work of F. Marin is financially supported by ANR funding (ACCRO-EARTH, ref. BLAN06-2_159971, coordinator Gilles Ramstein, LSCE) during the period 2007–2011.
Conflict of interest
None declared.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Marie, B., Joubert, C., Belliard, C. et al. Characterization of MRNP34, a novel methionine-rich nacre protein from the pearl oysters. Amino Acids 42, 2009–2017 (2012). https://doi.org/10.1007/s00726-011-0932-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00726-011-0932-0