Importance of The Location of The Negative-charged Counter-ion against The Protonated Schiff Base on The Chromophore Configuration of pharaonis Phoborhodopsin

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

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자료유형: 학술저널

저자정보: Shimono, Kazumi (Laboratory of Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University) Ikeura, Yukako (Laboratory of Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University) Sudo, Yuki (Laboratory of Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University) Iwamoto, Masayuki (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: 302 - 304 (3page)

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pharaonis phoborhodopsin (ppR), a photophobic sensor of haloalkaliphilic bacteria, Natronobacterium phar-aonis, has retinal as a chromophore covalently bound to Lys in G-helix via a protonated Schiff base (PSB), as is the same as bacteriorhodopsin (bR). For ppR, the corresponding counter-ion is Asp residue (Asp75) located in C-helix. Here we investigated the influence of the protonated state of this counter-ion and its location on the chromophore configuration. Under alkaline condition, the chromophore configuration of D75E mutant was analyzed by HPLC. D75E had a much larger content of 13-cis isomer: the ratio of 13-cis to all-trans was 6:4 while the wild-type had this ratio of 1 :9. On the other hand, under acidic condition where Glu was associated, D75E had no 13-cis retinal isomer. Mutants whose Asp75 was replaced by neutral amino acids (D75N and D75Q) did not contain 13-cis retinal. Furthermore, retinal isomer compositions and the change in the visible ab- sorption spectra (indicating the dissociation state of Glu75) were measured under varying pH, and these were almost the same dependencies. These results indicate that an important factor determining the 13-cis isomer content is the presence of negative charge of the counter-ion against PSB, but not the size of this residue. Com- parison between the wild-type and D75E in alkaline solutions indicates the influence of the location of the counter-ion.

#photosensor #archaea retinal protein #retinal isomer #counter-ion #protonated Schiff base

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