EXPRESSION, PURIFICATION, AND FUNCTIONAL ACTIVITY OF SHELL MATRIX PROTEIN PEARLIN FROM THE PEARL OYSTER PINCTADA FUCATA

文献类型： 外文期刊

第一作者： Huang, Guiju

作者： Huang, Guiju;Bi, Xiaomin;Zhang, Bo;Qu, Tao;Liu, Baosuo;Fan, Sigang;Yu, Dahui;Bi, Xiaomin

作者机构：

关键词： pearl oyster;Pinctada fucata;pearlin;MRC-5;MC3T3-E1;osteoblasts;proliferation;differentiation

期刊名称：JOURNAL OF SHELLFISH RESEARCH （ 影响因子：1.396； 五年影响因子：1.47 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Nacre, also known as mother of pearl, contains factors that can promote osteogenesis. In this study, pearlin, a protein from the mantle tissue of the pearl oyster Pinctada fucata, was expressed and purified, and its functional activity analyzed using two mineralogenic cell lines, MRC-5 (fibroblasts) and MC3T3-E1 (preosteoblasts). The open reading frame of pearlin was subcloned into the expression vector pET32a(+) and used to transform Escherichia coli BL21 (DE3) strain. The expression of the recombinant protein (molecular weight: 34.19 kDa) was induced by isopropyl-beta-D-thiogalactopyranoside in the form of inclusion bodies which were solubilized in 8 M urea. The recombinant protein was renatured by stepwise dialysis, purified by Ni-NTA affinity chromatography, and analyzed for the effects on cell proliferation by the MTT assay and osteoblastic differentiation by alkaline phosphatase (ALP) activity. The recombinant pearlin promoted proliferation of MRC-5 and MC3T3-E1 cells at 10 mu g/mL and increased their ALP activity at 2.5 mu g/mL and 5 mu g/mL, respectively. The present study showed that the recombinant pearlin exerted functional effects similar to those of the natural protein, laying a foundation for large-scale production of pearlin in a prokaryotic system.

分类号： Q7

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