小鼠Connexin31蛋白磷酸化定位  

Analysis of Mouse Connexin31 Phosphorylation Sites

作　　者：刘宇[1] 寻晓红[1] 

机构地区：[1]湖南理工学院化学化工系,湖南岳阳414006

出　　处：《湖南理工学院学报（自然科学版）》2009年第2期55-59,共5页Journal of Hunan Institute of Science and Technology(Natural Sciences)

基　　金：湖南省教育厅青年基金资助(07B029)

摘　　要：间隙连接蛋白31(Connexin31,Cx31)是间隙连接蛋白(Connexin)家族的一员,目前对于Cx31的功能及其调节方式知之甚少.本文通过免疫沉淀、SDS-PAGE分离、蛋白质条带回收、蛋白质胶块酶解、4-磺酸苯异硫氰酸酯修饰、PSD-MALDI-TOF质谱分析、数据分析、确定小鼠Cx31磷酸化位点.Connexin31（Cx31） is an important member of connexin 13 family. However, little is known about the Cx31 phosphorylation. In this study, we applied a phosphoproteomics approach to screen mouse Cx31 phosphorylation sites using HT1080 cells stable expressing a myc-tagged Cx31. The tryptic digests of proteins are chemically modified by 4-sulfophenyl isotlaiocyanate. The derivatization reaction introduces a negative sulfonic acid group at the N-terminus of a phosphopeptide, which can increase the efficiency of PSD fragmentation and enable the selective detection of only a single series of fragment ions（y-ions）. This chemical assisted method avoids the limitation of high background normally observed in MALDI-PSD spectra, and makes the spectra easier to interpret. In this study, serine 263 of the Cx31 was determined to be a site ofphosphorylation.

关 键 词：间隙连接蛋白31 免疫沉淀:磷酸化 

分 类 号：Q25[生物学—细胞生物学]