title: Crystal structure of Cruxrhodopsin-3 from Haloarcula vallismortis.
abstract: Cruxrhodopsin-3 (cR3), a retinylidene protein found in the claret membrane of Haloarcula vallismortis, functions as a light-driven proton pump. In this study, the membrane fusion method was applied to crystallize cR3 into a crystal belonging to space group P321. Diffraction data at 2.1 Å resolution show that cR3 forms a trimeric assembly with bacterioruberin bound to the crevice between neighboring subunits. Although the structure of the proton-release pathway is conserved among proton-pumping archaeal rhodopsins, cR3 possesses the following peculiar structural features: 1) The DE loop is long enough to interact with a neighboring subunit, strengthening the trimeric assembly; 2) Three positive charges are distributed at the cytoplasmic end of helix F, affecting the higher order structure of cR3; 3) The cytoplasmic vicinity of retinal is more rigid in cR3 than in bacteriorhodopsin, affecting the early reaction step in the proton-pumping cycle; 4) the cytoplasmic part of helix E is greatly bent, influencing the proton uptake process. Meanwhile, it was observed that the photobleaching of retinal, which scarcely occurred in the membrane state, became significant when the trimeric assembly of cR3 was dissociated into monomers in the presence of an excess amount of detergent. On the basis of these observations, we discuss structural factors affecting the photostabilities of ion-pumping rhodopsins.
authors.affiliation: Department of Physics, Graduate School of Science, Nagoya University, Nagoya, Japan.
authors.forename: Siu Kit
authors.initials: SK
authors.lastname: Chan
authors.affiliation: Department of Physics, Graduate School of Science, Nagoya University, Nagoya, Japan.
authors.forename: Tomomi
authors.initials: T
authors.lastname: Kitajima-Ihara
authors.affiliation: Department of Physics, Graduate School of Science, Nagoya University, Nagoya, Japan.
authors.forename: Ryudoh
authors.initials: R
authors.lastname: Fujii
authors.affiliation: Department of Physics, Graduate School of Science, Nagoya University, Nagoya, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Gotoh
authors.affiliation: Department of Physics, Graduate School of Science, Nagoya University, Nagoya, Japan.
authors.forename: Midori
authors.initials: M
authors.lastname: Murakami
authors.affiliation: Center for Gene Research, Nagoya University, Nagoya, Japan.
authors.forename: Kunio
authors.initials: K
authors.lastname: Ihara
authors.affiliation: Department of Physics, Graduate School of Science, Nagoya University, Nagoya, Japan; RIKEN Harima Institute/SPring-8, Mikazuki, Sayo, Hyogo, Japan.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Kouyama
mesh_terms: D000595:Amino Acid Sequence; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray; D015870:Gene Expression; D019613:Haloarcula / Q000737:chemistry*; D019617:Halobacterium salinarum / Q000235:genetics / Q000378:metabolism; D008027:Light; D008561:Membrane Fusion; D008969:Molecular Sequence Data; D055503:Protein Multimerization; D055550:Protein Stability; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry* / Q000235:genetics; D017494:Proton Pumps / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D025604:Rhodopsins, Microbial / Q000737:chemistry* / Q000235:genetics; D055672:Static Electricity; D014961:X-Ray Diffraction
chemical_list: D021122:Protein Subunits; D017494:Proton Pumps; D011994:Recombinant Proteins; D025604:Rhodopsins, Microbial