Localization of calmodulin and calmodulin-like protein and their functions in biomineralization in P. fucata
Localization of calmodulin and calmodulin-like protein and their functions in biomineralization in P. fucata
复制标题
钙调蛋白和钙调蛋白样蛋白的定位及其在 P. fucata 生物矿化中的功能
DOI：
10.1016/j.pnsc.2007.11.011
复制
发表时间：
2008-04-10
影响因子：
4.7
通讯作者：
Zhang, Rongqing
中科院分区：
文献类型：
--
作者：
Fang, Zi;Yan, Zhenguang;Zhang, Rongqing
钙调素（CaM）和钙调素样蛋白（卡尔普）是参与生物矿化的两种蛋白质。采用免疫印迹（WB）和免疫电镜技术研究了它们在合浦珠母贝外套膜上皮细胞中的定位。结果表明，这两种蛋白在亚细胞水平上的分布完全不同。CaM分布于外套膜上皮细胞的细胞核和细胞质中，而卡尔普仅分布于细胞质中。利用壳再生技术研究了这两种蛋白在生物矿化过程中的作用。在此过程中，外套膜上皮细胞内CaM和卡尔普的表达明显增强。这两种蛋白的过表达和在MC 3 T3-E1中缺乏额外的C-末端尾的钙调素样蛋白（M-CaLP）的突变体促进了成骨细胞的生物矿化标记物骨桥蛋白的mRNA表达。上述结果表明，CaM和卡尔普在外套膜上皮中的分布完全不同，并在不同水平上影响着生物矿化过程。卡尔普的C-末端额外的尾部对于其在合浦藻中的生物矿化中的功能是重要的。(C)2007年获国家自然科学基金、中国科学院院士。由Elsevier Limited和Science in China Press出版。All rights reserved.
Calmodulin (CaM) and calmodulin-like protein (CaLP) are two proteins involved in biomineralization. Their localizations in Pinctada fucata mantle epithelia were studied by Western blot (WB) analysis of the nuclear/cytosol fraction of primary cultured P. fucata mantle cells and immunogold electron microscopy. The results showed a completely different distribution of these two proteins at the subcellular level. CaM was distributed throughout both the nucleus and cytoplasm of the mantle epithelium but CaLP was distributed only in the cytoplasm. The functions of these two proteins in biomineralization were investigated by shell regeneration. During this process, the expressions of CaM and CaLP were greatly enhanced in different organelles of the mantle epithelium. Overexpression of these two proteins and a mutant of calmodulin-like protein (M-CaLP) that lacks an extra C-terminal tail in MC3T3-E1 promoted the mRNA expression of osteopontin, a biomineralization marker for osteoblasts. All of the results indicated that CaM and CaLP have completely different distributions in the mantle epithelium and affect the biomineralization process at different levels. The extra C-terminal tail of CaLP is important for its functions in biomineralization in P. fucata. (C) 2007 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.