Laser-induced transient grating analysis of dynamics of interaction between sensory rhodopsin II D75N and the HtrII transducer.
The interaction between sensory rhodopsin II (SRII) and its transducer HtrII was studied by the time-resolved laser-induced transient grating method using the D75N mutant of SRII, which exhibits minimal visible light absorption changes during its photocycle, but mediates normal phototaxis responses. Flash-induced transient absorption spectra of transducer-free D75N and D75N joined to 120 amino-acid residues of the N-terminal part of the SRII transducer protein HtrII (DeltaHtrII) showed only one spectrally distinct K-like intermediate in their photocycles, but the transient grating method resolved four intermediates (K(1)-K(4)) distinct in their volumes. D75N bound to HtrII exhibited one additional slower kinetic species, which persists after complete recovery of the initial state as assessed by absorption changes in the UV-visible region. The kinetics indicate a conformationally changed form of the transducer portion (designated Tr*), which persists after the photoreceptor returns to the unphotolyzed state. The largest conformational change in the DeltaHtrII portion was found to cause a DeltaHtrII-dependent increase in volume rising in 8 micros in the K(4) state and a drastic decrease in the diffusion coefficient (D) of K(4) relatively to those of the unphotolyzed state and Tr*. The magnitude of the decrease in D indicates a large structural change, presumably in the solvent-exposed HAMP domain of DeltaHtrII, where rearrangement of interacting molecules in the solvent would substantially change friction between the protein and the solvent.
Inoue
Keiichi
K
Department of Chemistry, Graduate School of Science, Kyoto University, Kyoto, Japan.
Sasaki
Jun
J
Spudich
John L
JL
Terazima
Masahide
M
D019843:Archaeal Proteins / Q000502:physiology* / Q000528:radiation effects*; D002338:Carotenoids / Q000502:physiology* / Q000528:radiation effects*; D004307:Dose-Response Relationship, Radiation; D007700:Kinetics; D007834:Lasers; D020283:Photoreceptors, Microbial / Q000502:physiology* / Q000528:radiation effects; D011829:Radiation Dosage; D015398:Signal Transduction / Q000502:physiology* / Q000528:radiation effects
D019843:Archaeal Proteins; C434451:HtrII protein, Natronobacterium pharaonis; D020283:Photoreceptors, Microbial; C434450:phototaxis receptor sensory rhodopsin II, Natronobacterium pharaonis; D002338:Carotenoids