title: Dynamics of light-induced conformational changes of the phoborhodopsin/transducer complex formed in the n-dodecyl beta-D-maltoside micelle.
abstract: A complex of photoreceptor phoborhodopsin (ppR; also called sensory rhodopsin II) and its cognate halobacterial transducer II (pHtrII) existing in the plasma membrane mediates the light signal to the cytoplasm in the earliest step of negative phototaxis in Natronomonas pharaonis. We have investigated the dynamics of the light-induced conformational changes of the ppR/pHtrII(1-159) complex formed in the presence of 0.1% n-dodecyl beta-d-maltoside (DDM) by a fluorescence resonance energy transfer (FRET) based method. Fluorescence donor and acceptor dyes were linked to cysteine residues genetically introduced at given positions in pHtrII and ppR. The light-induced FRET efficiency changes for various pairs of dye-labeled cysteine residues were determined to examine dynamics of movements of given residues in the transmembrane and the linker region including the HAMP domain in pHtrII induced by photoexcitation of ppR. Upon flash excitation of ppR, FRET efficiency changed depending on pairs of the labeled cysteine residues. The distances between V185 in ppR and the five given residues (102 through 141) in the pHtrII linker region estimated from the FRET efficiency increased by 0.3-0.8 A; on the other hand, the distances between S31 in ppR and the five residues in pHtrII decreased. The changes arose within 70 ms (the dead time of instrument) and decayed at a rate of 1.1 +/- 0.2 s. Azide significantly increased the decay rate of light-induced FRET efficiency changes by accelerating the decay of the M state of ppR. The decay rate of FRET efficiency changes coincided with the rate of recovery of the ppR to the initial state but not the decay of the M state. We conclude that the light-induced conformational change of pHtrII occurs before, at the formation or during the M state, and its relaxation is coupled tightly with the decay of the O state of ppR in the 1:1 complex formed in the DDM micelle.
authors: Nano-biotechnology Research Center and Department of Bioscience, School of Science and Technology, Kwansei Gakuin University, Sanda, Hyogo, Japan.
authors: Yukinori
authors: Y
authors: Taniguchi
authors: Tatsuya
authors: T
authors: Ikehara
authors: Naoki
authors: N
authors: Kamo
authors: Hiroshi
authors: H
authors: Yamasaki
authors: Yoshinori
authors: Y
authors: Toyoshima
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry* / Q000378:metabolism*; D003902:Detergents; D031541:Fluorescence Resonance Energy Transfer; D005960:Glucosides / Q000737:chemistry / Q000378:metabolism*; D025602:Halorhodopsins / Q000737:chemistry* / Q000378:metabolism*; D008027:Light; D008823:Micelles; D010782:Photolysis; D011487:Protein Conformation; D025603:Sensory Rhodopsins / Q000737:chemistry* / Q000378:metabolism*; D013816:Thermodynamics; D014785:Vision, Ocular
chemical_list: D019843:Archaeal Proteins; D003902:Detergents; D005960:Glucosides; D025602:Halorhodopsins; D008823:Micelles; D025603:Sensory Rhodopsins; C061508:sensory rhodopsin II protein, archaeal; C040358:dodecyl maltoside