Proteomic analysis of the organic matrix of the abalone Haliotis asinina calcified shell

BackgroundThe formation of the molluscan shell is regulated to a large extent by a matrix of extracellular macromolecules that are secreted by the shell forming tissue, the mantle. This so called "calcifying matrix" is a complex mixture of proteins and glycoproteins that is assembled and occluded within the mineral phase during the calcification process. While the importance of the calcifying matrix to shell formation has long been appreciated, most of its protein components remain uncharacterised.ResultsRecent expressed sequence tag (EST) investigations of the mantle tissue from the tropical abalone (Haliotis asinina) provide an opportunity to further characterise the proteins in the shell by a proteomic approach. In this study, we have identified a total of 14 proteins from distinct calcified layers of the shell. Only two of these proteins have been previously characterised from abalone shells. Among the novel proteins are several glutamine- and methionine-rich motifs and hydrophobic glycine-, alanine- and acidic aspartate-rich domains. In addition, two of the new proteins contained Kunitz-like and WAP (whey acidic protein) protease inhibitor domains.ConclusionThis is one of the first comprehensive proteomic study of a molluscan shell, and should provide a platform for further characterization of matrix protein functions and interactions.

[1]  M. Mann,et al.  In-depth, high-accuracy proteomics of sea urchin tooth organic matrix , 2008 .

[2]  J. Pais de Barros,et al.  The shell matrix of the freshwater mussel Unio pictorum (Paleoheterodonta, Unionoida) , 2007, The FEBS journal.

[3]  M. Fritz,et al.  Perlwapin, an abalone nacre protein with three four-disulfide core (whey acidic protein) domains, inhibits the growth of calcium carbonate crystals. , 2006, Biophysical journal.

[4]  Takeshi Takeuchi,et al.  Biphasic and Dually Coordinated Expression of the Genes Encoding Major Shell Matrix Proteins in the Pearl Oyster Pinctada fucata , 2005, Marine Biotechnology.

[5]  E. Dalas,et al.  The effect of aminoacids on the crystal growth of calcium carbonate , 2002 .

[6]  B. Ganss,et al.  Calcium binding to sialic acids and its effect on the conformation of ependymins. , 1993, European journal of biochemistry.

[7]  B. Marie,et al.  Evolution of Nacre: Biochemistry and Proteomics of the Shell Organic Matrix of the Cephalopod Nautilus macromphalus , 2009, Chembiochem : a European journal of chemical biology.

[8]  Arul Marie,et al.  Heterogeneity of Proteinase Inhibitors in the Water-Soluble Organic Matrix from the Oyster Nacre , 2007, Marine Biotechnology.

[9]  T Morita,et al.  A carbonic anhydrase from the nacreous layer in oyster pearls. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[10]  M. Fritz,et al.  Perlustrin, a Haliotis laevigata (abalone) nacre protein, is homologous to the insulin-like growth factor binding protein N-terminal module of vertebrates. , 2001, Biochemical and biophysical research communications.

[11]  M. Fritz,et al.  Perlinhibin, a cysteine-, histidine-, and arginine-rich miniprotein from abalone (Haliotis laevigata) nacre, inhibits in vitro calcium carbonate crystallization. , 2007, Biophysical journal.

[12]  Steve Weiner,et al.  Mollusk shell formation: a source of new concepts for understanding biomineralization processes. , 2006, Chemistry.

[13]  Takashi Kato,et al.  An Acidic Matrix Protein, Pif, Is a Key Macromolecule for Nacre Formation , 2009, Science.

[14]  I. SlnasurNA,et al.  Primary structure of a soluble matrix protein of scallop shell: Implications for calcium carbonate biomineralization , 1998 .

[15]  Michael Kube,et al.  Parallel evolution of nacre building gene sets in molluscs. , 2010, Molecular biology and evolution.

[16]  B. Degnan,et al.  Dynamic expression of ancient and novel molluscan shell genes during ecological transitions , 2007, BMC Evolutionary Biology.

[17]  V. Shashoua,et al.  Evidence for the in vivo polymerization of ependymin: a brain extracellular glycoprotein , 1990, Brain Research.

[18]  Gert Wörheide,et al.  A rapidly evolving secretome builds and patterns a sea shell , 2006, BMC Biology.

[19]  Xiao-yan Wang,et al.  Identification and characterization of a biomineralization related gene PFMG1 highly expressed in the mantle of Pinctada fucata. , 2007, Biochemistry.

[20]  M. Mann,et al.  The sea urchin (Strongylocentrotus purpuratus) test and spine proteomes , 2008, Proteome Science.

[21]  T. Fujikawa,et al.  Structures of mollusc shell framework proteins , 1997, Nature.

[22]  P. Hansma,et al.  Molecular Cloning and Characterization of Lustrin A, a Matrix Protein from Shell and Pearl Nacre of Haliotis rufescens * , 1997, The Journal of Biological Chemistry.

[23]  Takuma Nishida,et al.  Expression of genes responsible for biomineralization of Pinctada fucata during development. , 2010, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[24]  J. Evans,et al.  Characterization of two molluscan crystal‐modulating biomineralization proteins and identification of putative mineral binding domains , 2003, Biopolymers.

[25]  A. Poustka,et al.  Proteomic analysis of sea urchin (Strongylocentrotus purpuratus) spicule matrix , 2010, Proteome Science.

[26]  Y. Yamakoshi Dentinogenesis and Dentin Sialophosphoprotein (DSPP). , 2009, Journal of oral biosciences.

[27]  Feng Ding,et al.  The Length Dependence of the PolyQ-mediated Protein Aggregation* , 2007, Journal of Biological Chemistry.

[28]  M. Kirkland,et al.  MERP1: a mammalian ependymin-related protein gene differentially expressed in hematopoietic cells. , 2002, Gene.

[29]  Jesper V Olsen,et al.  Proteomic analysis of the acid‐soluble organic matrix of the chicken calcified eggshell layer , 2006, Proteomics.

[30]  M. Fritz,et al.  The amino-acid sequence of the abalone (Haliotis laevigata) nacre protein perlucin. Detection of a functional C-type lectin domain with galactose/mannose specificity. , 2000, European journal of biochemistry.

[31]  Evolution of large body size in abalones (Haliotis): patterns and implications , 2005, Paleobiology.

[32]  Benjamin Marie,et al.  Molluscan shell proteins: primary structure, origin, and evolution. , 2008, Current topics in developmental biology.