Structural basis for the slow photocycle and late proton release in Acetabularia rhodopsin I from the marine plant Acetabularia acetabulum

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• 作者：Munenori Furuse ; Jun Tamogami ; Toshiaki Hosaka ; Takashi Kikukawa ; Naoko Shinya ; Masakatsu Hato ; Noboru Ohsawa ; So Young Kim ; Kwang-Hwan Jung ; Makoto Demura ; Seiji Miyauchi ; Naoki Kamo ; Kazumi Shimono ; Tomomi Kimura-Someya ; Shigeyuki Yokoyama and Mikako Shirouzu
• 关键词：light-driven ion pump ; cell-free protein synthesis ; X-ray crystal structure ; microbial rhodopsin ; membrane protein
• 刊名：Acta Crystallographica Section D
• 出版年：2015
• 出版时间：November 2015
• 年：2015
• 卷：71
• 期：11
• 页码：2203-2216
• 全文大小：2846K
• ISSN：1399-0047

文摘

Although many crystal structures of microbial rhodopsins have been solved, those with sufficient resolution to identify the functional water molecules are very limited. In this study, the Acetabularia rhodopsin I (ARI) protein derived from the marine alga A. acetabulum was synthesized on a large scale by the Escherichia coli cell-free membrane-protein production method, and crystal structures of ARI were determined at the second highest (1.52–1.80 Å) resolution for a microbial rhodopsin, following bacteriorhodopsin (BR). Examinations of the photochemical properties of ARI revealed that the photocycle of ARI is slower than that of BR and that its proton-transfer reactions are different from those of BR. In the present structures, a large cavity containing numerous water molecules exists on the extracellular side of ARI, explaining the relatively low pK_a of Glu206^ARI, which cannot function as an initial proton-releasing residue at any pH. An interhelical hydrogen bond exists between Leu97^ARI and Tyr221^ARI on the cytoplasmic side, which facilitates the slow photocycle and regulates the pK_a of Asp100^ARI, a potential proton donor to the Schiff base, in the dark state.