Photoreaction pathways and photointermediates of retinal-binding photoreceptor proteins as revealed by in situ photoirradiation solid-state NMR spectroscopy
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Photoirradiation solid-state NMR spectroscopy is a powerful means to study photoreceptor retinal-binding proteins by the detection of short-lived photointermediates to elucidate the photoreaction cycle and photoactivated structural changes. An in situ photoirradiation solid-state NMR apparatus has been developed for the irradiation of samples with extremely high efficiency to enable observation of photointermediates which are stationary trapped states. Such observation enables elucidation of the photoreaction processes of photoreceptor membrane proteins. Therefore, in situ photoirradiation is particularly useful study the photocycle of retinal-binding proteins such as sensory rhodopsin I (SRI) and sensory rhodopsin II (SRII) because functional photointermediates have relatively longer half-lives than other photointermediates. As a result, several photointermediates have been trapped as stationary state and their detailed structures and photoreaction cycles have been revealed using photoirradiation solid-state NMR spectroscopy at low temperature. Photoreaction intermediates of bacteriorhodopsin, which functions to provide light-driven proton pump activity, were difficult to trap because the half-lives of the photointermediates were shorter than those of sensory rhodopsin. Therefore, these photointermediates are trapped in a freeze-trapped state at a very low temperature and the NMR signals were observed using a combination of photoirradiation and dynamic nuclear polarization (DNP) experiments.
KeywordsSensory rhodopsin Bacteriorhodopsin Photoreaction cycle Photointermediate Photoirradiation solid-state NMR
This work was supported by the Grants-in-Aid for Scientific Research in an Innovative Area (JP16H00756 to AN and JP16H00828 to IK), and by the Grants-in-Aid for Scientific Research (C) (JP15K06963 to AN) and Research (B) (JP18H02387 to IK) from the Japan Society for the Promotion of Science (JSPS).
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Conflict of interest
Akira Naito declares that he has no conflict of interest. Yoshiteru Makino declares that he has no conflict of interest. Arisu Shigeta declares that she has no conflict of interest. Izuru Kawamura declares that he has no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
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