期刊
JOURNAL OF PROTEOME RESEARCH
卷 16, 期 6, 页码 2282-2293出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.7b00154
关键词
EGFR; laser photolysis; hydroxyl radical; footprinting; mass spectrometry
资金
- Singapore Ministry of Education (Tier 2) [ARC9/15]
- National Medical Research Council of Singapore [NMRC-OF-IRG-0003-2016]
- NTU-NHG Ageing Research Grant [ARG/14017]
Mass spectrometry-based protein footprinting emerged as a useful technology to understand protein ligand interactions in vitro. We have previously demonstrated the application of footprinting in live E. coli cells. Here, we further optimized an ultrafast laser photolysis hydroxyl radical footprinting method and applied, it to study the interaction of EGF and EGFR in live mammalian cells. This method used a nanosecond laser to photochemically generate a burst of hydroxyl radicals in situ in-cell suspension to oxidize the amino acids on the protein surface. Mass spectrometric analysis of the thus modified peptides was interpreted to probe the solvent-accessible surface areas of the protein in its native biological state with and without EGF activation. Our footprinting data agreed with the two relevant EGFR crystal structures, indicating, that this in-cell laser photolysis footprinting technique is a valid approach to study the structural properties of integral membrane proteins directly in the native environment.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据