嗜铬颗粒蛋白B与热休克同源蛋白70相互作用的研究

首都医科大学学报 ›› 2005, Vol. 26 ›› Issue (6): 699-702.

嗜铬颗粒蛋白B与热休克同源蛋白70相互作用的研究

油红捷, 王泽生

首都医科大学基础医学院生物化学与分子生物学系

收稿日期:2004-10-09 修回日期:1900-01-01 出版日期:2005-12-24 发布日期:2005-12-24
通讯作者: 王泽生

Study of the Interaction between Chromogranin B and Heat Shock Cognate Protein70

You Hongjie, Wang Zesheng

Department of Biochemistry and Molecular Biology, School of Basic Medicine, Capital University of Medical Sciences

Received:2004-10-09 Revised:1900-01-01 Online:2005-12-24 Published:2005-12-24

摘要/Abstract

摘要：

目的分离和鉴别在肾上腺髓质嗜铬颗粒细胞分泌囊泡中,与嗜铬颗粒蛋白B(CgB)相互作用的蛋白质。方法采用蛋白质捕获、免疫共沉淀和质谱等技术进行研究。结果在嗜铬颗粒细胞内,CgB与热休克同源蛋白70(HSC 170)相互作用。一个HSC 170结合序列定位在CgB的羧基末端区(Ser586-Arg602,SR-17),而且SR-17内的Ser595磷酸化可增强其与HSC 170结合。磷酸化修饰的SR-17可从ATP-Sepharose上洗脱HSC 170。用免疫印迹和免疫组化方法可证实CgB和HSC 170均存在于嗜铬颗粒细胞的胞质和细胞核中。结论 HSC 170可能参与CgB的细胞核转位过程。

关键词: 嗜铬颗粒蛋白B, 热休克同源蛋白70, 磷酸化修饰, 细胞核转位

Abstract:

Objective To investigate the role that HSC 170 plays in the process of CgBtranslocation in chromaffin cells.Methods Pull-down, co-immunoprecipitation and mass spectrometry of proteins were used to identify the proteins interested.Results The present data show an interaction of CgBwith heat shock cognate protein 70(Hsc70) in the chromaffin cell.Abinding sequence of Hsc70 is determined in the C-terminal region of porcine CgB(Ser586-Arg602, SR-17).Moreover, phosphorylation of Ser595 in pCgBenhances its association with Hsc70.On the other hand, Hsc70 is eluted from ATP-Sepharose by incubation with phosphorylated SR-17 at Ser595.Western blot analyses show that both CgBand Hsc70 are present in the nucleus and cytoplasm of these cells.Conclusion The present results suggest that Hsc70 may participate in the process of CgBtranslocation in chromaffin cells and that phosphorylation of CgBis likely to mediate this process.

Key words: chromogranin B, heat shock cognate protein 70, phosphorylation, nuclear translocation

中图分类号:

油红捷;王泽生. 嗜铬颗粒蛋白B与热休克同源蛋白70相互作用的研究[J]. 首都医科大学学报, 2005, 26(6): 699-702.

You Hongjie;Wang Zesheng. Study of the Interaction between Chromogranin B and Heat Shock Cognate Protein70[J]. Journal of Capital Medical University, 2005, 26(6): 699-702.