글로벌 연구동향
분자영상 및 방사화학
- 2016년 12월호
[J Am Chem Soc.] Observing Extremely Weak Protein-Protein Interactions with Conventional Single-Molecule Fluorescence Microscopy.연세대/ 유장현, 윤태영*
- 출처
- J Am Chem Soc.
- 등재일
- 2016 Nov 2
- 저널이슈번호
- 138(43):14238-14241. Epub 2016 Oct 25.
- 내용
Abstract
Extremely weak protein-protein interactions (PPIs), signified by micromolar or even millimolar dissociation constants, are one of the keys to understanding the rapid responses of cellular systems. Although single-molecule methods are particularly useful in determining kinetics of biological processes, their application is largely limited to rather strong interactions because of the diffraction-limited observation volume. In this study, we report a single-molecule method that allows the characterization of PPIs using a prey concentration 4 orders of magnitude lower than the dissociation constant. Instead of increasing the concentration of diffusing molecules, which is inevitably limited by the optical diffraction limit, we employed an increased density of surface bait protein. The low occupancy of the surface baits permitted determination of the kinetics with single-molecule resolution. We used this approach to study a PPI network consisting of Ras and its downstream proteins including full-length Rafs and catalytic subunits of phosphoinositide 3-kinase.
Author information
Yoo J1,2,3, Lee TS2,3, Choi B1,2,3, Shon MJ2,3, Yoon TY2,3.
1Department of Physics, Korea Advanced Institute of Science and Technology (KAIST) , Daejeon 34141, South Korea.
2Center for Nanomedicine, Institute for Basic Science (IBS), Yonsei University , Seoul 30722, South Korea.
3Yonsei-IBS Institute, Yonsei University , Seoul 30722, South Korea.
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