Identification of new centrosome proteins by autoimmune patient sera  

作　　者：XIA Liang1, LI Yan1, YANG Dong1, WANG LiMin1, HE Fang1, ZHOU ChunYuan1, LI YongZhe2, ZENG ChangQing1 & He DaCheng1 1 Key Laboratory for Cell Proliferation and Regulation Biology of Ministry of Education, Beijing Normal University, Beijing 100875, China 2 Peking Union Medical Hospital, Peking Union Medical College, Beijing 100730, China 3 Beijing Genomic Institute, Beijing 101300, China 

出　　处：《Science China(Life Sciences)》2007年第2期194-202,共9页中国科学（生命科学英文版）

基　　金：the special fundations of Ministry of Education of China and National Natural Science Foundation of China (Grant No. 30225017)

摘　　要：Compared to other subcellular organelles, centrosome proteome can hardly be studied, due to the dif- ficulties in separation and purification of centrosome. Auto-antisera from 6 autoimmune patients, which recognized centrosome specifically in immunofluorescence, were used to identify the corresponding centrosomal proteins. The sera were first tested by Western blot on whole cell lysate, and all bound antibodies were then eluted from each single band in Western blot membrane to assure which antibody was responsible for the centrosome specific immunofluorescence staining. The corresponding pro- teins were obtained by immunoprecipitation and identified by mass spectrometry. Six centrosomal proteins, including 2 known centrosomal proteins and 4 proteins with unknown localization or report- edly non-centrosomal localization, were identified. These proteins apparently involve in cell cycle regulation, signal transduction pathways, molecular chaperons, and metabolism enzymes, which may reflect the expected functional diversity of centrosome.Compared to other subcellular organelles, centrosome proteome can hardly be studied, due to the dif- ficulties in separation and purification of centrosome. Auto-antisera from 6 autoimmune patients, which recognized centrosome specifically in immunofluorescence, were used to identify the corresponding centrosomal proteins. The sera were first tested by Western blot on whole cell lysate, and all bound antibodies were then eluted from each single band in Western blot membrane to assure which antibody was responsible for the centrosome specific immunofluorescence staining. The corresponding pro- teins were obtained by immunoprecipitation and identified by mass spectrometry. Six centrosomal proteins, including 2 known centrosomal proteins and 4 proteins with unknown localization or report- edly non-centrosomal localization, were identified. These proteins apparently involve in cell cycle regulation, signal transduction pathways, molecular chaperons, and metabolism enzymes, which may reflect the expected functional diversity of centrosome.

关 键 词：CENTROSOME AUTOIMMUNE PATIENT SERA ELUTION of antibody immunoflourescence IMMUNOPRECIPITATION mass spectrometry 

分 类 号：R593.2[医药卫生—内科学]