Asp83, Glu113 and Glu134 are not specifically involved in Schiff base protonation or wavelength regulation in bovine rhodopsin

FEBS Letters

De Caluwé, G.L.J.¹  De Grip, W.J.¹  Janssen, J.J.M.¹ 

[1] Department of Biochemistry, University of Nijmegen, PO Box 9101, 6500 HB Nijmegen, The Netherlands

关键词: Amino acid substitution;    Baculovirus;    Heterologous expression;    Rhodopsin;    Spectral property;    Site selective mutagenesis;    AcNPV;    Autographa californica nuclear polyhedrosis virus;    bp;    base pair;    dpi;    days post-infection;    MOI;    multiplicity of infection;    CHAPS;    3-[(3-cholamidopropyl)dimethylammonio]propanosulfonate;    DTE;    1;    4-dithioerythritol;    EDTA;    (ethylene-1;    2-diamine)N-tetraacetic acid;    Con A;    concanavalin A;    G-protein;    GTP-binding protein or transducin;    RT;    room temperature;    v-ops;    opsin produced in vitro by recombinant baculovirus;    v-rho;    rhodopsin obtained by incubation of v-ops with 11-Z-retinal;   

DOI  :  10.1016/0014-5793(90)80080-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Site-specific mutagenesis was employed to investigate the proposed contribution of proton-donating residues (Glu, Asp) in the membrane domains of bovine rhodopsin to protonation of the Schiff base-linking protein and chromophore or to wavelength modulation of this visual pigment. Three point-mutations were introduced to replace the highly conserved residues Asp₈₃ by Asn (D₈₃N), Glu₁₁₃ by Gln (E₁₁₃Q) or Glu₁₃₄ by Asp (E₁₃₄D), respectively. All 3 substitutions had only marginal effects on the spectral properties of the final pigment (⩽ 3 nm blue-shift relative to native rhodopsin). Hence, none of these residues by itself is specifically involved in Schiff base protonation or wavelength modulation of bovine rhodopsin.

【 授权许可】

Unknown