The mollusc shell is a hard tissue consisting of calcium carbonate and organic matrices. The organic matrices are believed to play important roles in shell formation. In the present study, we extracted and purified a novel matrix protein, named Prismalin-14, from the acid-insoluble fraction of the prismatic layer of the shell of the Japanese pearl oyster (Pinctada fucata), and determined its whole amino acid sequence by a combination of amino acid sequence analysis and MS analysis of the intact protein and its enzymic digests. Prismalin-14 consisted of 105 amino acid residues, including PIYR repeats, a Gly/Tyr-rich region and N- and C-terminal Asp-rich regions. Prismalin-14 showed inhibitory activity on calcium carbonate precipitation and calcium-binding activity in vitro. The scanning electron microscopy images revealed that Prismalin-14 affected the crystallization of calcium carbonate in vitro. A cDNA encoding Prismalin-14 was cloned and its expression was analysed. The amino acid sequence deduced from the nucleotide sequence of Prismalin-14 cDNA was identical with that determined by peptide sequencing. Northern-blot analysis showed that a Prismalin-14 mRNA was expressed only at the mantle edge. In situ hybridization demonstrated that a Prismalin-14 mRNA was expressed strongly in the inner side of the outer fold of the mantle. These results suggest that Prismalin-14 is a framework protein that plays an important role in the regulation of calcification of the prismatic layer of the shell.
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Research Article|
August 10 2004
Characterization of Prismalin-14, a novel matrix protein from the prismatic layer of the Japanese pearl oyster (Pinctada fucata)
Michio SUZUKI;
Michio SUZUKI
*Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, Yayoi 1-1-1, Bunkyo, Tokyo 113-8657, Japan
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Emi MURAYAMA;
Emi MURAYAMA
*Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, Yayoi 1-1-1, Bunkyo, Tokyo 113-8657, Japan
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Hirotaka INOUE;
Hirotaka INOUE
*Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, Yayoi 1-1-1, Bunkyo, Tokyo 113-8657, Japan
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Noriaki OZAKI;
Noriaki OZAKI
†Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Kawaguchi 4-1-8, Saitama 332-0012, Japan
‡Department of Earth and Planetary Science, Graduate School of Science, University of Tokyo, Hongo 7-3-1, Bunkyo, Tokyo 113-0033, Japan
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Hidekazu TOHSE;
Hidekazu TOHSE
*Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, Yayoi 1-1-1, Bunkyo, Tokyo 113-8657, Japan
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Toshihiro KOGURE;
Toshihiro KOGURE
‡Department of Earth and Planetary Science, Graduate School of Science, University of Tokyo, Hongo 7-3-1, Bunkyo, Tokyo 113-0033, Japan
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Hiromichi NAGASAWA
Hiromichi NAGASAWA
1
*Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, Yayoi 1-1-1, Bunkyo, Tokyo 113-8657, Japan
1To whom correspondence should be addressed (email anagahi@mail.ecc.u-tokyo.ac.jp).
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Publisher: Portland Press Ltd
Received:
March 01 2004
Revision Received:
April 19 2004
Accepted:
May 07 2004
Accepted Manuscript online:
May 07 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2004
Biochem J (2004) 382 (1): 205–213.
Article history
Received:
March 01 2004
Revision Received:
April 19 2004
Accepted:
May 07 2004
Accepted Manuscript online:
May 07 2004
Citation
Michio SUZUKI, Emi MURAYAMA, Hirotaka INOUE, Noriaki OZAKI, Hidekazu TOHSE, Toshihiro KOGURE, Hiromichi NAGASAWA; Characterization of Prismalin-14, a novel matrix protein from the prismatic layer of the Japanese pearl oyster (Pinctada fucata). Biochem J 15 August 2004; 382 (1): 205–213. doi: https://doi.org/10.1042/BJ20040319
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