Influence of Arg72 of pharaonis Phoborhodopsin on M-intermediate Decay and Proton Pumping Activity

Ikeura, Yukako; Shimono, Kazumi; Iwamoto, Masayuki; Sudo, Yuki; Kamo, Naoki

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자료유형: 학술저널

저자정보: Ikeura, Yukako (Laboratory of Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University) Shimono, Kazumi (Laboratory of Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University) Iwamoto, Masayuki (Laboratory of Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University) Sudo, Yuki (Laboratory of Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University) Kamo, Naoki (Laboratory of Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University)

저널정보: 한국광과학회 Journal of photoscience : an international journal officail organ of the korean society of photoscience Journal of photoscience : an international journal officail organ of the korean society of photoscience 제9권 제2호

발행연도: 2002.1

수록면: 311 - 313 (3page)

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X-ray structures of pharaonis phoborhodopsin (ppR) show the different direction of the side chain of Arg72 from that of the corresponding residue (Arg82) of bacteriorhodopsin, BR. For BR, this residue is considered to play an important role in the proton pumping. In order to investigate the role of Arg72 in ppR, we constructed Arg72 mutants of R72A, R72K and R72Q, and measured the photocycle and proton pumping activities. The pH-titration curves on the absorption maximum of the mutants were shifted to alkaline in comparison of that of the wild-type. This may imply the increase of pKa of D75, suggesting the presence of the (probably electric) interaction between D75 and Arg72. Rate constants of the M-decay were 3-7 times faster than that of the wild-type, and the time for the completion of the photocycling was also reduced. Using Sn0$_2$ electrode, the rate of transmembrane proton transport was measured upon illumination. The photo-induced proton pumping activities were estimated after the corrections that are the percentages of the associated form of D75 (which has no pumping activity) and the photocycling rates. R72A and R72Q showed the reduced activity while R72K did not reduce the activity.

#Super-conserved Arg #Photocycling #pKa of the counterion of the Schiff base #Hydrogen bonding network #$Sn0_2$ electrode #Archaeal rhodopsin #Sensory rhodopsin

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