纳升电喷雾串联质谱技术对阻断泛素通路诱导凋亡相关蛋白质的鉴定  

Nano ESI MS/MS Identification on Apoptosis Associated Proteins Induced by Inhibiting Ubiquitin proteasome Pathway

作　　者：王红霞[1] 靳宝锋[2] 王杰[1] 何昆[1] 杨松成[1] 沈倍奋[2] 张学敏[1] 

机构地区：[1]军事医学科学院国家生物医学分析中心,北京100850 [2]军事医学科学院基础医学研究所,北京100850

出　　处：《生物化学与生物物理学报》2002年第5期630-634,共5页

基　　金：国家高技术研究发展计划 ( 86 3计划 ) (No .2 0 0 1AA2 3 3 0 41);国家自然科学基金资助项目 (No .3 0 0 70 3 77)

摘　　要：应用毛细管液相色谱 电喷雾 四极杆 飞行时间串联质谱和纳升电喷雾 四极杆 飞行时间串联质谱技术 ,对阻断白血病细胞泛素通路诱发的凋亡相关蛋白质进行了鉴定。通过双向电泳发现 ,蛋白质斑点H在阻断Mo7e白血病细胞泛素通路 2h之后 ,表达量明显增加 ,6h达到最高。该斑点经MALDI TOF MS肽质量指纹谱分析未获结果 ,但通过上述 2种串联质谱技术获得其胰蛋白酶水解肽段的串联质谱图和肽段的全长序列 ,经检索均确认为RhoGDIβ蛋白。进一步发现阻断泛素通路还诱发了另 2个斑点出现 ,位于H点附近 ,经鉴定为同一蛋白质 。CapLC ESI MS/MS and nano ESI MS/MS techniques were used to identify the apoptosis associated proteins induced by inhibiting the ubiquitin proteasome pathway in Mo7e leukaemic cells. In 2 DE, spot H was found to initiate its overexpression at 2 h after the inhibition and reached its peak at 6 h. It was identified as Rho GDI β protein after the tandem mass spectrum and after the sequence of its tryptic peptides were obtained by the ESI MS/MS techniques. It was not revealed by peptide mass fingerprint using MALDI TOF MS. Other two spots induced by the inhibition appeared close to spot H were also revealed identical to Rho GDI β, possibly due to unknown modifications.

关 键 词：纳升电喷雾串联质谱技术 阻断 泛素通路 诱导 凋亡相关蛋白质 鉴定 

分 类 号：Q51[生物学—生物化学] Q503