期刊
EXPERT REVIEW OF PROTEOMICS
卷 14, 期 7, 页码 617-626出版社
TAYLOR & FRANCIS LTD
DOI: 10.1080/14789450.2017.1345632
关键词
Chromatin; cross-talk; histones; mass spectrometry; middle-down; post-translational modifications; proteomics
资金
- U.S. Department of Health and Human Services, National Institute of Health [R01GM110174, P01CA196539]
- U.S. Department of Defense [W81XWH-113-1-0426]
- Leukemia and Lymphoma Society
Introduction: Analysis of histone post-translational modifications (PTMs) by mass spectrometry (MS) has become a fundamental tool for the characterization of chromatin composition and dynamics. Histone PTMs benchmark several biological states of chromatin, including regions of active enhancers, active/repressed gene promoters and damaged DNA. These complex regulatory mechanisms are often defined by combinatorial histone PTMs; for instance, active enhancers are commonly occupied by both marks H3K4me1 and H3K27ac. The traditional bottom-up MS strategy identifies and quantifies short (aa 4-20) tryptic peptides, and it is thus not suitable for the characterization of combinatorial PTMs.Areas covered: Here, we review the advancement of the middle-down MS strategy applied to histones, which consists in the analysis of intact histone N-terminal tails (aa 50-60). Middle-down MS has reached sufficient robustness and reliability, and it is far less technically challenging than PTM quantification on intact histones (top-down). However, the very few chromatin biology studies applying middle-down MS resulting from PubMed searches indicate that it is still very scarcely exploited, potentially due to the apparent high complexity of method and analysis.Expert commentary: We will discuss the state-of-the-art workflow and examples of existing studies, aiming to highlight its potential and feasibility for studies of cell biologists interested in chromatin and epigenetics.
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