报告题目：Label-free single-molecule quantification of chemically
报 告 人：Jie Yan教授，新加坡国立大学
Chemically induced dimerization (CID) refers to utilizing small molecules to induce dimerization of two protein domains. CID has been applied to study numerous biological processes through precise spatiotemporal control of protein-protein interactions. In addition, CID also has important pharmaceutical applications. CID involves complex multi-step interactions. Precise quantification of each step of the interactions is critical to our understanding of the stability and kinetics of the final ternary complex formed through CID. CID is often associated with very slow off rates in some of the steps, making them technically challenging to be quantified using most of the current biochemistry assays. In addition, in many applications the final ternary complex is subject to various mechanical constraints, which cannot be quantified using current technologies. Here, I will present a highly efficient label-free single-molecule assay that can quantify each step of CID reactions and demonstrate the application of the technology by quantifying each key interaction involved in rapamycin induced heterodimerization of FRB and FKBP protein domains. I will also show that the high mechanical stability of the FRB-rapamycin-FKBP ternary complex can be utilized to control mechanosensing of cells.
Yan Jie is currently a professor at Department of Physics at National University of Singapore. He is also a principal investigator at NUS Mechanobiology Institute and NUS Centre for BioImaging Sciences. In 2015, Yan Jie was awarded American Physical Society (APS) Fellowship. And in 2016, he received the Singapore National Research Foundation (NRF) Investigatorship as well as the Human Frontier Science Program’s Research Grants. He has published 99 research papers on journals including Nature, Nature Communication, PNAS, and eLife. He received one PhD in experimental biophysics form University of Illinois, Chicago (2005), and one PhD from Institute of Theoretical Physics, Chinese Academy of Sciences (1998).