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APEX, a new genetically encoded tag for spatial proteome mapping & EM imaging

 

2013-03-12

attach file : file포스터_031213.pptx

Hyun-Woo Rhee, Ph.D (UNIST)

Seminar / 2013 March 12th 04:00pm / EB1-711

Abstract

Microscopy and mass spectrometry (MS) are complementary techniques: the former provides spatiotemporal information in living cells, but only for a handful of recombinant proteins, while the latter can detect thousands of endogenous proteins simultaneously, but only in lysed samples. In this session, I will introduce a new technology that combines these strengths by offering spatially- and temporally-resolved proteomic maps of endogenous proteins within living cells. The method relies on a genetically-targetable peroxidase enzyme (APEX) that biotinylates nearby proteins promiscuously, which are subsequently purified and identified by MS. Our recent proteomics mapping result of mitochondrial matrix using APEX-generated phenoxyl radical biotinylation in living cells will be discussed. The radical generation and polymerization activity of APEX could be also useful to produce distinct electron-microscopy (EM) contrast to achieve EM-level identification of proteins of interest simultaneously.