Proton Release and Uptake of pharaonis Phoborhodopsin (Sensory Rhodopsin II) Reconstituted into Phospholipids

Masayuki Iwamoto, Chisa Hasegawa, Yuki Sudo, Kazumi Shimono, Tsunehisa Araiso, Naoki Kamo

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

pharaonis phoborhodopsin (ppR, also called pharaonis sensory rhodopsin II, psRII) is a photo-receptor for negative phototaxis in Natronobacterium pharaonis. During the photoreaction cycle (photocycle), ppR exhibits intraprotein proton movements, resulting in proton pumping from the cytoplasmic to the extracellular side, although it is weak. In this study, light-induced proton uptake and release of ppR reconstituted with phospholipid were analyzed using a SnO2 electrode. The reconstituted ppR exhibited properties in proton uptake and release that are different from those of dodecyl maltoside solubilized samples. It showed fast proton release before the decay of ppR M (M-photointermediate) followed by proton uptake, which was similar to that of bacteriorhodopsin (BR), a light-driven proton pump. Mutant analysis assigned Asp193 to one (major) of the members of the proton-releasing group (PRG). Fast proton release was observed only when the pH was approximately 5-8 in the presence of Cl-. When Cl- was replaced with SO 42-, the reconstituted ppR did not exhibit fast proton release at any pH, suggesting Cl- binding around PRG. PRG in BR consists of Glu204 (Asp193 in ppR) and Glu194 (Pro183 in ppR). Replacement of Pro183 by Glu/Asp, a negatively charged residue, led to Cl --independent fast proton release. The transducer binding affected the properties of PRG in ppR in the ground state and in the ppRM state, suggesting that interaction with the transducer extends to the extracellular surface of ppR. Differences and similarities in the molecular mechanism of the proton movement between ppR and BR are discussed.

Original languageEnglish
Pages (from-to)3195-3203
Number of pages9
JournalBiochemistry
Volume43
Issue number11
DOIs
Publication statusPublished - Mar 23 2004
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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