Metabolic labeling in middle-down proteomics allows for investigation of the dynamics of the histone code.

ABSTRACT:

Background

Middle-down mass spectrometry (MS), i.e., analysis of long (~50-60 aa) polypeptides, has become the method with the highest throughput and accuracy for the characterization of combinatorial histone posttranslational modifications (PTMs). The discovery of histone readers with multiple domains, and overall the cross talk of PTMs that decorate histone proteins, has revealed that histone marks have synergistic roles in modulating enzyme recruitment and subsequent chromatin activities. Here, we demonstrate that the middle-down MS strategy can be combined with metabolic labeling for enhanced quantification of histone proteins and their combinatorial PTMs in a dynamic manner.

Methods

We used a nanoHPLC-MS/MS system consisting of hybrid weak cation exchange-hydrophilic in

SUBMITTER: Sidoli S 

PROVIDER: S-EPMC5501349 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature