TAILIEUCHUNG - Báo cáo khoa học: The structure–function relationship analysis of Prismalin-14 from the prismatic layer of the Japanese pearl oyster, Pinctada fucata

The mollusk shell is a hard tissue consisting of calcium carbonate and organic matrices. The organic matrices are considered to play important roles in shell formation. We have previously identified a prismatic layer-specific protein named Prismalin-14, which consists of 105 amino acid resi-dues and includes four structurally characteristic regions; | ễFEBS Journal The structure-function relationship analysis of Prismalin-14 from the prismatic layer of the Japanese pearl oyster Pinctada fucata Michio Suzuki and Hiromichi Nagasawa Department of Applied BiologicalChemistry Graduate Schoolof Agriculturaland Life Sciences University of Tokyo Japan Keywords biomineralization chitin pearl oyster Pinctada fucata Prismalin-14 Correspondence H. Nagasawa Department of Applied BiologicalChemistry Graduate Schoolof Agriculturaland Life Sciences University of Tokyo 1-1-1 Yayoi Bunkyo Tokyo 1138657 Japan Fax 81 3 5841 8022 Tel 81 3 5841 5132 E-mail anagahi@ Received 5 July 2007 revised 7 August 2007 accepted 8 August 2007 doi The mollusk shell is a hard tissue consisting of calcium carbonate and organic matrices. The organic matrices are considered to play important roles in shell formation. We have previously identified a prismatic layerspecific protein named Prismalin-14 which consists of 105 amino acid residues and includes four structurally characteristic regions a repeated sequence of Pro-Ile-Tyr-Arg a Gly Tyr-rich region and N- and C-terminal Asp-rich regions. Prismalin-14 showed an inhibitory activity on calcium carbonate precipitation and a calcium-binding ability in vitro. In this study we prepared some molecular species of recombinant proteins including Prismalin-14 and its truncated proteins in an Escherichia coli expression system to reveal a structure-function relationship of Prismalin-14. The results showed that the Gly Tyr-rich region was responsible for chitin binding and was identified as a novel chitin-binding sequence. On the other hand both N- and C-terminal Asp-rich regions are related to inhibitory activity on calcium carbonate precipitation in vitro. Immunohistological observation revealed that Prismalin-14 was localized at the acid-insoluble organic framework including chitin. All these results strongly suggest that Prismalin-14 is a framework .

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