著者

Marie Kurihara Vera Thiel Hirona Takahashi Keiichi Kojima David M. Ward Donald A. Bryant Makoto Sakai Susumu Yoshizawa Yuki Sudo

出版者

The Pharmaceutical Society of Japan

雑誌

Chemical and Pharmaceutical Bulletin (ISSN:00092363)

巻号頁・発行日

vol.71, no.2, pp.154-164, 2023-02-01 (Released:2023-02-01)

参考文献数

45

被引用文献数

1

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Rhodopsins are transmembrane proteins with retinal chromophores that are involved in photo-energy conversion and photo-signal transduction in diverse organisms. In this study, we newly identified and characterized a rhodopsin from a thermophilic bacterium, Bellilinea sp. Recombinant Escherichia coli cells expressing the rhodopsin showed light-induced alkalization of the medium only in the presence of sodium ions (Na+), and the alkalization signal was enhanced by addition of a protonophore, indicating an outward Na+ pump function across the cellular membrane. Thus, we named the protein Bellilinea Na+-pumping rhodopsin, BeNaR. Of note, its Na+-pumping activity is significantly greater than that of the known Na+-pumping rhodopsin, KR2. We further characterized its photochemical properties as follows: (i) Visible spectroscopy and HPLC revealed that BeNaR has an absorption maximum at 524 nm with predominantly (>96%) the all-trans retinal conformer. (ii) Time-dependent thermal denaturation experiments revealed that BeNaR showed high thermal stability. (iii) The time-resolved flash-photolysis in the nanosecond to millisecond time domains revealed the presence of four kinetically distinctive photointermediates, K, L, M and O. (iv) Mutational analysis revealed that Asp101, which acts as a counterion, and Asp230 around the retinal were essential for the Na+-pumping activity. From the results, we propose a model for the outward Na+-pumping mechanism of BeNaR. The efficient Na+-pumping activity of BeNaR and its high stability make it a useful model both for ion transporters and optogenetics tools.

著者

坂井 誠 Makoto SAKAI

雑誌

恵泉女学園大学紀要 = Keisen University Bulletin

巻号頁・発行日

no.27, pp.57-75, 2015-02

著者

酒井 誠 Makoto Sakai

雑誌

中京大学大学院生法学研究論集 = Chukyo Daigaku daigakuinsei hogaku kenkyu ronshu (ISSN:03897958)

巻号頁・発行日

vol.1, pp.1-49, 1981-03-01

著者

坂井 誠 サカイ マコト Makoto SAKAI

雑誌

恵泉女学園大学紀要 = Keisen University Bulletin

巻号頁・発行日

vol.20, pp.131-158, 2008-02