日本質量分析学会 第66回質量分析総合討論会

プログラム

ポスター発表

第2日 5月16日(水)  ポスター会場

SWATH-MSによる‘chromatome’の網羅的解析

(1東大院理2九大生医研)
o幡野敦1松本雅記2中山敬一2黒田真也1

Chromatin is a complex of DNA and proteins which forms chromosomes within the nucleus of eukaryotic cells. The primary functions of chromatin are packaging DNA into a more compact shape, controlling gene expression and DNA replication. These biological processes are selectively mediated by the different set of proteins under particular conditions or stimuli. Therefore comprehensive identification of chromatin-associated proteins ‘chromatome’ provides useful information to understand how cells respond to extracellular stimuli via chromatin reorganization. Recently, Kustatscher et al. developed Chromatin Enrichment For Proteomics (ChEP), which is a simple biochemical procedure that enriches interphase chromatin in all its complexity.
The aim of our study is elucidation of mechanistic regulation of various functions of chromatin at systems-wide scale via a mathematical model, which strongly demands accurate quantitative data on various cultural conditions. We set up SWATH-MS analysis, a data independent acquisition (DIA) method, which achieves high-throughput, high-reproducibility and consistency of identification and quantification of proteins, to characterize ‘chromatome’ of cells stimulated with various condition. As a proof of concept demonstration, we performed ChEP of hepatoma cells stimulated with insulin in the presence of kinase inhibitors by using SWATH-MS. These analyses provided quantitative data of chromatin-associated proteins responding to insulin at proteome-wide scale.