Quantitative comparison of protein dynamics in live cells and in vitro by in-cell 19F-NMR

Yousuke Takaoka(Kyoto Katsura Hospital), Yoshiyuki Kioi(Kyoto University), Akira Morito(Kyoto University), Junji Otani(Kyoto University), Kyohei Arita(Kyoto University), Eishi Ashihara(Kyoto Pharmaceutical University), Mariko Ariyoshi(Kyoto University), Hidehito Tochio(Kyoto University), Masahiro Shirakawa(Kyoto University), Itaru Hamachi(Japan Science and Technology Agency)
Chemical Communications
January 1, 2013
10.1039/c3cc39205h
Cited by 56

Abstract

Here we describe how a (19)F-probe incorporated into an endogenous protein by a chemical biology method revealed protein dynamics. By explicit determination of ligand-bound and unbound structures with X-ray crystallography, the quantitative comparison of the protein's dynamics in live cells and in vitro is presented. These results clearly demonstrated the greater conformational fluctuations of the intracellular protein, partially due to macromolecular crowding effects.

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