In-Situ Photo Irradiation Solid-State NMR Spectroscopy Applied to Retinal-Binding Membrane Proteins

  • Akira NaitoEmail author
  • Yoshiteru MakinoEmail author
  • Izuru KawamuraEmail author
Living reference work entry


It is essential to elucidate the photoreaction cycles to understand the function of photoreceptor membrane proteins. The photointermediates that appear in the photoreaction cycle typically have short half-lives; therefore, it is difficult to detect the photointermediates using solid-state nuclear magnetic resonance (NMR) spectroscopy. Recently, it has become possible to observe photointermediates using in situ photoirradiation solid-state NMR and dynamic nuclear polarization (DNP)-enhanced photoirradiation solid-state NMR. In situ photoirradiation solid-state NMR is designed to irradiate light from the top part of the zirconia rotor through a glass cap, which makes it possible to irradiate the inside of the rotor and leads to efficient photoirradiation. This method has made it possible to observe photointermediates of sensory rhodopsins such as sensory rhodopsin I (SRI) and sensory rhodopsin II (SRII), and bacteriorhodopsin (bR) and its Y185F mutant. In SRI, green light generates M-intermediates that show positive phototaxis, while blue light generates P-intermediates that show negative phototaxis. In SRII, green light generates M-intermediates and blue light generates O-intermediates. In Y185F-bR, O-intermediate was first observed using solid-state NMR spectroscopy. DNP-enhanced photoirradiation is also a powerful tool for the detection of short-lived photointermediate such as K-, L-, M-, and N-intermediates in bR. In channel rhodopsin, several photointermediates such as P1, P2, P3, P4, and Px photointermediates have been observed. Thus, in situ photoirradiation solid-state NMR spectroscopy is demonstrated to provide significant insights to understand the photoactivation mechanisms of photoreceptor retinal-binding membrane proteins.


Photoreaction cycle Photoreceptor membrane protein Photointermediate In situ photoirradiation Solid-state NMR Dynamic nuclear polarization Rhodopsin Sensory rhodopsin Bacteriorhodopsin Channel rhodopsin Phototaxis 


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© Springer International Publishing AG 2017

Authors and Affiliations

  1. 1.Graduate School of EngineeringYokohama National UniversityHodogaya-kuJapan

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