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Article

Photoreaction Pathways of Bacteriorhodopsin and Its D96N Mutant as Revealed by in Situ Photoirradiation Solid-State NMR

1
Graduate School of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama 240-8501, Japan
2
Laboratory of Organic Chemistry for Life Science, Kobe Pharmaceutical University, 4-19-1 Motoyamakitamachi, Higashinada-ku, Kobe 658-8558, Japan
*
Authors to whom correspondence should be addressed.
Academic Editor: Mingxu You
Membranes 2022, 12(3), 279; https://doi.org/10.3390/membranes12030279
Received: 28 December 2021 / Revised: 25 February 2022 / Accepted: 26 February 2022 / Published: 28 February 2022
Bacteriorhodopsin (BR) functions as a light-driven proton pump that transitions between different states during the photocycle, such as all-trans (AT; BR568) and 13-cis, 15-syn (CS; BR548) state and K, L, M1, M2, N, and O intermediates. In this study, we used in situ photoirradiation 13C solid-state NMR to observe a variety of photo-intermediates and photoreaction pathways in [20-13C]retinal-WT-BR and its mutant [20-13C, 14-13C]retinal-D96N-BR. In WT-BR, the CS state converted to the CS* intermediate under photoirradiation with green light at −20 °C and consequently converted to the AT state in the dark. The AT state converted to the N intermediate under irradiation with green light. In D96N-BR, the CS state was converted to the CS* intermediate at −30 °C and consequently converted to the AT state. Simultaneously, the AT state converted to the M and L intermediates under green light illumination at −30 °C and subsequently converted to the AT state in the dark. The M intermediate was directly excited to the AT state by UV light illumination. We demonstrated that short-lived photo-intermediates could be observed in a stationary state using in situ photoirradiation solid-state NMR spectroscopy for WT-BR and D96N-BR, enabling insight into the light-driven proton pump activity of BR. View Full-Text
Keywords: bacteriorhodopsin; light driven proton pump; photo-intermediate; photoirradiation solid-state NMR bacteriorhodopsin; light driven proton pump; photo-intermediate; photoirradiation solid-state NMR
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MDPI and ACS Style

Shigeta, A.; Otani, Y.; Miyasa, R.; Makino, Y.; Kawamura, I.; Okitsu, T.; Wada, A.; Naito, A. Photoreaction Pathways of Bacteriorhodopsin and Its D96N Mutant as Revealed by in Situ Photoirradiation Solid-State NMR. Membranes 2022, 12, 279. https://doi.org/10.3390/membranes12030279

AMA Style

Shigeta A, Otani Y, Miyasa R, Makino Y, Kawamura I, Okitsu T, Wada A, Naito A. Photoreaction Pathways of Bacteriorhodopsin and Its D96N Mutant as Revealed by in Situ Photoirradiation Solid-State NMR. Membranes. 2022; 12(3):279. https://doi.org/10.3390/membranes12030279

Chicago/Turabian Style

Shigeta, Arisu, Yuto Otani, Ryota Miyasa, Yoshiteru Makino, Izuru Kawamura, Takashi Okitsu, Akimori Wada, and Akira Naito. 2022. "Photoreaction Pathways of Bacteriorhodopsin and Its D96N Mutant as Revealed by in Situ Photoirradiation Solid-State NMR" Membranes 12, no. 3: 279. https://doi.org/10.3390/membranes12030279

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