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Photoreaction Pathways of Bacteriorhodopsin and Its D96N Mutant as Revealed by in Situ Photoirradiation Solid-State NMR
| Content Provider | MDPI |
|---|---|
| Author | Shigeta, Arisu Otani, Yuto Miyasa, Ryota Makino, Yoshiteru Kawamura, Izuru Okitsu, Takashi Wada, Akimori Naito, Akira |
| Copyright Year | 2022 |
| Description | 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, $M_{1}$, $M_{2}$, N, and O intermediates. In this study, we used in situ photoirradiation$ ^{13}$C solid-state NMR to observe a variety of photo-intermediates and photoreaction pathways in $[20-^{13}$C]retinal-WT-BR and its mutant $[20-^{13}$C, $14-^{13}$C]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. |
| Starting Page | 279 |
| e-ISSN | 20770375 |
| DOI | 10.3390/membranes12030279 |
| Journal | Membranes |
| Issue Number | 3 |
| Volume Number | 12 |
| Language | English |
| Publisher | MDPI |
| Publisher Date | 2022-02-28 |
| Access Restriction | Open |
| Subject Keyword | Membranes Biochemistry and Molecular Biology Bacteriorhodopsin Light Driven Proton Pump Photo-intermediate Photoirradiation Solid-state Nmr |
| Content Type | Text |
| Resource Type | Article |
