title: Protein-protein interaction of a Pharaonis halorhodopsin mutant forming a complex with Pharaonis halobacterial transducer protein II detected by Fourier-transform infrared spectroscopy.
abstract: Pharaonis halorhodopsin (pHR) functions as a light-driven inward chloride ion pump in Natoronomonas pharaonis, while pharaonis phoborhodopsin (ppR; also called pharaonis sensory rhodopsin II, pSRII), is a light sensor for negative phototaxis. ppR forms a 2:2 complex with its cognate transducer protein (pHtrII) through intramembranous hydrogen bonds: Tyr199(ppR)-Asn74(pHtrII) and Thr189(ppR)-Glu43 (pHtrII), Ser62(pHtrII). It was reported that a pHR mutant (P240T/F250Y), which possesses the hydrogen-bonding sites, impairs its pumping activity upon complexation with pHtrII. In this study, effect of the complexation with pHtrII on the structural changes upon formation of the K, L(1) and L(2) intermediates of pHR was investigated by use of Fourier-transform infrared spectroscopy. The vibrational changes of Tyr250(pHR) and Asn74(pHtrII) were detected for the L(1) and L(2) intermediates, supporting that Tyr250(pHR) forms a hydrogen bond with Asn74(pHtrII) as similarly to Tyr199(ppR). The conformational changes of the retinal chromophore were never affected by complexation with pHtrII, but amide-I vibrations were clearly different in the absence and presence of pHtrII. The molecular environment around Asp156(pHR) in helix D is also slightly affected. These additional structural changes are probably related to blocking of translocation of a chloride ion from the extracellular to the cytoplasmic side during the photocycle.
authors.affiliation: Department of Materials Science and Engineering, Nagoya Institute of Technology, Showa-ku, Nagoya, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Furutani
authors.forename: Motohiro
authors.initials: M
authors.lastname: Ito
authors.forename: Yuki
authors.initials: Y
authors.lastname: Sudo
authors.forename: Naoki
authors.initials: N
authors.lastname: Kamo
authors.forename: Hideki
authors.initials: H
authors.lastname: Kandori
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D025602:Halorhodopsins / Q000737:chemistry*; D006860:Hydrogen Bonding; D008027:Light; D019633:Natronobacterium / Q000737:chemistry*; D025603:Sensory Rhodopsins / Q000737:chemistry*; D013055:Spectrophotometry, Infrared; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods
chemical_list: D000596:Amino Acids; D025602:Halorhodopsins; D025603:Sensory Rhodopsins; C061508:sensory rhodopsin II protein, archaeal