Analysis of nuclear lipidation in gene regulation

点击翻译按钮获取中文摘要

This year we performed proteomic analysis of purified endogenous HN4 protein using the scheme established last year.First of all, we purified endogenous HNF4 protein by affinity purification, then samples were subjected to LC-MS/MS analysis. From this experiment, we identified 14 PTM sites in HNF4 protein such as phosphorylation, acetylation, methylation and ubiquitilation. This result was reported to the scientific journal as an original paper(Yokoyama et al., Biochem Biophys Res Commun, 2011, 410(4): 749-5). However, we could not identify the lipidation of HNF4 in this experiment.Furthermore, we adopted the novel method "click-it reaction" for the proteomic analysis of nuclear lipidation. In this method, labeled proteins were purified by avidin beads, and then subjected to the LC-MS/MS analysis. As a result, we identified various proteins including HNF4 and histone protein. We think this result implies that nuclear lipidation has more global role than we expected before, and now we are preparing the manuscript for the paper concerning this results.