ホソイ　ハルコ   Hosoi Haruko   細井　晴子    所属   東邦大学  理学部 生物分子科学科    職種   准教授
論文種別	原著
言語種別	英語
査読の有無	査読あり
表題	Three Simultaneous Fluorescence Resonance Energy Transfer Processes and Structural Relaxation of Enhanced Yellow Fluorescent Protein Observed by Picosecond Time-Resolved Fluorescence Anisotropy
掲載誌名	正式名：The Journal of Physical Chemistry B 略　 称：J. Phys. Chem. B ISSNコード：15206106/1520-5207
掲載区分	国外
出版社	American Chemical Society
巻・号・頁	125(29),pp.7997-8009
総ページ数	13
著者・共著者	H. Tsubota, A. Takayama, Y. Takeda, N. Yamada, H. Hosoi
担当区分	最終著者,責任著者
発行年月	2021/06/14
概要	Fluorescent proteins (FPs) have been widely used to visualize biological processes in living cells. It is essential to understand the underlying fluorescence mechanism to develop novel FPs and to interpret imaging data appropriately. Enhanced yellow fluorescent protein (eYFP) is one of the most typical FPs; however, several reports to date have been limited to individual discussion, which is insufficient to understand the full picture of the dynamics involved. In this study, we focused on the fluorescence resonance energy transfer (FRET) and dimerization behavior and performed picosecond time-resolved fluorescence measurements of eYFP and its A206K mutant, which does not form a dimer. The combination of the dissociation constant and the acid dissociation constant rationally explains the mechanism of ultrafast homo-FRET and ultrafast hetero-FRET. It is also shown that structural relaxation occurs in the dimer after excited-state proton transfer. The formation efficiencies and quaternary structures of dimers consisting of different protonation states are shown to be different. Furthermore, under high-concentration conditions, “slow” homo-FRET with tens of nanoseconds timescale occurs between monomers and dimers. The findings from this study will be applied to other fluorescent proteins such as Aequorea victoria green FP and its mutants and various red FPs with longer conjugation lengths.