文献类型：期刊文献

英文题名：Visualization of the pH-dependent dynamic distribution of G2A in living cells

作者：Lan, Wanjun[1,4];Yamaguchi, Satoshi[1];Yamamoto, Teruyasu[1];Yamahira, Shinya[2];Tan, Modong[1];Murakami, Naoka[3];Zhang, Jingyan[4];Nakamura, Motonao[3];Nagamune, Teruyuki[1,2]

机构：[1]Univ Tokyo, Dept Chem & Biotechnol, Tokyo 1138656, Japan;[2]Univ Tokyo, Grad Sch Engn, Dept Bioengn, Tokyo 1138656, Japan;[3]Univ Tokyo, Grad Sch Med, Dept Biochem & Mol Biol, Tokyo 1138656, Japan;[4]E China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab New Drug Design, State Key Lab Bioreactor Engn, Shanghai, Peoples R China

年份：2014

卷号：28

期号：9

起止页码：3965

外文期刊名：FASEB JOURNAL

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000340845900014)】；

基金：This work was supported by a Ministry of Education, Culture, Sports, Science and Technology, Japan, Grant-in-Aid for Challenging Exploratory Research 246564498 and a grant for the Center for NanoBio Integration (CNBI). W. L. was supported by the China Scholarship Council (CSC) and the National Science Foundation of China (No. 31070742).

语种：英文

外文关键词：GPCR; internalization; pH dependence; pulse labeling

摘要：G2A (from G2 accumulation) receptor is a member of the proton-sensing G-protein coupled receptor (GPCR) family and induces signal transduction events that regulate the cell cycle, proliferation, oncogenesis, and immunity. The mechanism by which G2A-mediated signal transduction is regulated by the extracellular pH remains unresolved. Here, we first visualize the pH-dependent G2A distribution change in living cells by a sortase A-mediated pulse labeling technology: the short-peptide tag-fused human G2A on human embryo kidney HEK293T cell surfaces was labeled with a small fluorescent dye in the presence of lysophosphatidylcholine, and the labeled G2A was chased at acidic and neutral pHs in real time by microscope time course observations. G2A internalization from cell surfaces into intracellular compartments was observed to be inhibited under acidic pH conditions, and this inhibition was relieved at neutral pH. Additionally, the internalized G2A was redistributed onto cell surfaces by jumping from a neutral to an acidic pH. From quantitative image analysis data, we conclude the amount of G2A on the cell surface was controlled by suppressing the G2A internalization rate by one-tenth in response to the extracellular acidic pH, and this acidic pH-induced G2A accumulation on cell surfaces may be explained by proton-induced dissociation of G2A from endocytic machinery.