Siuti Nertila, Roth Michael J, Mizzen Craig A, Kelleher Neil L, Pesavento James J
Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
J Proteome Res. 2006 Feb;5(2):233-9. doi: 10.1021/pr050268v.
The basis set of protein forms expressed by human cells from the H2B gene family was determined by Top Down Mass Spectrometry. Using Electron Capture Dissociation for MS/MS of H2B isoforms, direct evidence for the expression of unmodified H2B.Q, H2B.A, H2B.K/T, H2B.J, H2B.E, H2B.B, H2B.F, and monoacetylated H2B.A was obtained from asynchronous HeLa cells. H2B.A was the most abundant form, with the overall expression profile not changing significantly in cells arrested in mitosis by colchicine or during mid-S, mid-G2, G2/M, and mid-G1 phases of the cell cycle. Modest hyperacetylation of H2B family members was observed after sodium butyrate treatment.
通过自上而下的质谱分析法确定了人类细胞中由H2B基因家族表达的蛋白质形式的基础集。使用电子捕获解离对H2B异构体进行串联质谱分析,从异步培养的HeLa细胞中获得了未修饰的H2B.Q、H2B.A、H2B.K/T、H2B.J、H2B.E、H2B.B、H2B.F和单乙酰化H2B.A表达的直接证据。H2B.A是最丰富的形式,在秋水仙碱使细胞停滞于有丝分裂期或细胞周期的S中期、G2中期、G2/M期和G1中期时,其整体表达谱没有显著变化。丁酸钠处理后观察到H2B家族成员有适度的超乙酰化。