한빛사 논문
Jiwoong Kwon1,9, Jong-Seok Park2,8,9, Minsu Kang1,3, Soobin Choi4, Jumi Park5, Gyeong Tae Kim6, Changwook Lee5, Sangwon Cha4, Hyun-Woo Rhee2,7,* & Sang-Hee Shim1,3,*
1 Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science (IBS), Seoul 02841, Republic of Korea.
2 Department of Chemistry, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.
3 Department of Chemistry, Korea University, Seoul 02841, Republic of Korea.
4 Department of Chemistry, Hankuk University of Foreign Studies, Yongin 17035, Republic of Korea.
5 Department of Biological Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.
6 Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.
7 Department of Chemistry, Seoul National University, Seoul 08826, Republic of Korea.
8 Present address: SK Biopharmaceuticals Co., Ltd.,, Daejeon 34124, Republic of Korea.
9 These authors contributed equally: Jiwoong Kwon, Jong-Seok Park.
*Correspondence to Hyun-Woo Rhee or Sang-Hee Shim
Abstract
We introduce UnaG as a green-to-dark photoswitching fluorescent protein capable of high-quality super-resolution imaging with photon numbers equivalent to the brightest photoswitchable red protein. UnaG only fluoresces upon binding of a fluorogenic metabolite, bilirubin, enabling UV-free reversible photoswitching with easily controllable kinetics and low background under Epi illumination. The on- and off-switching rates are controlled by the concentration of the ligand and the excitation light intensity, respectively, where the dissolved oxygen also promotes the off-switching. The photo-oxidation reaction mechanism of bilirubin in UnaG suggests that the lack of ligand-protein covalent bond allows the oxidized ligand to detach from the protein, emptying the binding cavity for rebinding to a fresh ligand molecule. We demonstrate super-resolution single-molecule localization imaging of various subcellular structures genetically encoded with UnaG, which enables facile labeling and simultaneous multicolor imaging of live cells. UnaG has the promise of becoming a default protein for high-performance super-resolution imaging.
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TOP52020년 후보
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