Characterization of Prismalin-14, a novel matrix protein from the prismatic layer of the Japanese pearl oyster (Pinctada fucata)
Characterization of Prismalin-14, a novel matrix protein from the prismatic layer of the Japanese pearl oyster (Pinctada fucata)
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DOI：
10.1042/bj20040319
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发表时间：
2004-08-15
影响因子：
4.1
通讯作者：
Nagasawa, H
中科院分区：
文献类型：
--
作者：
Suzuki, M;Murayama, E;Nagasawa, H
软体动物的壳是一种由碳酸钙和有机基质组成的硬组织。有机基质被认为在壳的形成中起重要作用。本研究从合浦珠母贝（Pinctada fucata）壳棱柱层酸不溶性组分中提取纯化了一种新的基质蛋白，命名为Prismalin-14，并通过氨基酸序列分析和质谱分析确定了其完整蛋白及其酶活性的氨基酸序列。Prismalin-14由105个氨基酸残基组成，包括PIYR重复序列、Gly/Tyr富集区以及N-和C-末端Asp富集区。Prismalin-14在体外表现出对碳酸钙沉淀和钙结合活性的抑制活性。扫描电子显微镜图像显示Prismalin-14在体外影响碳酸钙的结晶。克隆了编码Prismalin-14的cDNA并分析了其表达。从Prismalin-14 cDNA的核苷酸序列推导的氨基酸序列与肽序列测定的氨基酸序列一致。Northern印迹分析表明，Prismalin-14 mRNA仅在外套膜边缘表达。原位杂交结果表明，Prismalin-14 mRNA强烈表达于外套膜外褶的内侧。这些结果表明，Prismalin-14是一种框架蛋白，在调节壳的棱柱层的钙化中起着重要作用。
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.