Date : March 8, 2018 16:30 ~
Speaker : Prof. Hyun-Woo Rhee(SNU)
Location : Magam Hall, Bldg 500
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Proximity labeling is an in situ protein biotinylation reaction by enzymatically generated reactive
biotin-species which have a short lifetime (less than 1 ms) in aqueous solution. Recently, proximity
labeling has shown remarkable new findings in biological research fields such as sub-organelle
proteome mapping and in vivo interactome identification which have not been easily obtained by
conventional methods. Our group recently developed an improved proximity labeling method (Spot-ID)
by mass-spec detection of the biotin-labeled proteins which are generated by engineered ascorbate
peroxidase (APEX) or promiscuous biotin ligase (pBirA). This method allows the direct identification
of the labeled protein without false positive findings and the structural identification of the labeled
protein in live cells. Using this method, we could map in vivo proteomic architecture of the inner
mitochondrial membrane (IMM) whose membrane topology has not been fully characterized yet.
Furthermore, this method allowed the identification of new elements in the rapamycin-induced
interactome on the FK506-rapamycin binding (FRB) domain of mTOR in living cells. In this seminar,
I will introduce a new approach to perform proximity crosslinking by utilization of a simple coupling
reaction in live cells. This method could identify physically interacting molecular partner(s) of protein
of interest in live cells. Overall, we could observe that these new methods are very useful to unveil
systems biology in live cells.