Asp 83, Glu 113 and Glu 134 are not specifically involved in Schiff base protonation or wavelength regulation in bovine rhodopsin

• Published in: FEBS letters Vol. 260; no. 1; pp. 113 - 118
• Main Authors: Janssen, J.J.M., De Caluwé, G.L.J., De Grip, W.J.
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 15.01.1990
• Subjects: Amino acid substitution; Baculovirus; Heterologous expression; Rhodopsin; Site selective mutagenesis; Spectral property; Rhodopsin; Spectral property; v-rho, rhodopsin obtained by incubation of v-ops with 11-Z-retinal; AcNPV, Autographa californica nuclear polyhedrosis virus; Baculovirus; CHAPS, 3-[(3-cholamidopropyl)dimethylammonio]propanosulfonate; v-ops, opsin produced in vitro by recombinant baculovirus; Amino acid substitution; Con A, concanavalin A; MOI, multiplicity of infection; EDTA, (ethylene-1,2-diamine) N-tetraacetic acid; bp, base pair; DTE, 1,4-dithioerythritol; dpi, days post-infection; Heterologous expression; Site selective mutagenesis; G-protein, GTP-binding protein or transducin; RT, room temperature
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(90)80080-3

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 Asp 83 by Asn (D 83N), Glu 113 by Gln (E 113Q) or Glu 134 by Asp (E 134D), 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.