Protein mass analysis of histones

被引:31
作者
Galasinski, SC
Resing, KA
Ahn, NG [1 ]
机构
[1] Univ Colorado, Dept Chem & Biochem, Boulder, CO 80309 USA
[2] Univ Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USA
关键词
histories; high-mobility-group proteins; modifications; mass spectrometry; chromatin; acetylation; methylation; phosphorylation;
D O I
10.1016/S1046-2023(03)00082-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Posttranslational modification of chromatin-associated proteins, including histories and high-mobility-group (HMG) proteins, provides an important mechanism to control gene expression, genome integrity, and epigenetic inheritance. Protein mass analysis provides a rapid and unbiased approach to monitor multiple chemical modifications on individual molecules. This review describes methods for acid extraction of histories and HMG proteins, followed by separation by reverse-phase chromatography coupled to electrospray ionization mass spectrometry (LC/ESI-MS). Posuranslational modifications are detected by analysis of full-length protein masses. Confirmation of protein identity and modification state is obtained through enzymatic digestion and peptide sequencing by MS/MS. For differentially modified forms of each protein, the measured intensities are semiquantitative and allow determination of relative abundance and stoichiometry. The method simultaneously detects covalent modifications oil Multiple proteins and provides a facile assay for comparing chromatin modification states between different cell types and/or cellular responses. (C) 2003 Elsevier Science (USA). All rights reserved.
引用
收藏
页码:3 / 11
页数:9
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