title: Characterization of rhodopsin congenital night blindness mutant T94I.
abstract: The Thr94 --> Ile mutation in the second transmembrane segment of rhodopsin has been reported to be associated with a congenital night blindness phenotype in a large Irish pedigree. Previously, two other known rhodopsin mutants that cause congenital night blindness, A292E and G90D, have been shown in vitro to constitutively activate the G protein transducin in the absence of a chromophore. The proposed mechanism of constitutive activation of these two mutants is an electrostatic disruption of the active site salt bridge between Glu113 and Lys296 that contributes to stabilization of the protein in the inactive state. Here, the T94I rhodopsin mutant is characterized and compared to the two other known rhodopsin night blindness mutants. The T94I mutant opsin is shown also to constitutively activate transducin. The T94I mutant pigment (with a bound 11-cis-retinal chromophore), like the other known rhodopsin night blindness mutants, is not active in the dark and has wild-type activity upon exposure to light. Similar to the Gly90 --> Asp substitution, position 94 is close enough to the Schiff base nitrogen that an Asp at this position can functionally substitute for the Glu113 counterion. However, in contrast to the other night blindness mutants, the T94I MII intermediate decays with a half-life that is approximately 8-fold slower than in the wild-type MII intermediate. Thus, the one phenotype shared by all congenital night blindness mutants that is different from the wild-type protein is constitutive activation of the apoprotein.
authors.affiliation: Department of Biochemistry, Brandeis University, Waltham, Massachusetts 02454, USA.
authors.forename: Alecia K
authors.initials: AK
authors.lastname: Gross
authors.forename: Vikram R
authors.initials: VR
authors.lastname: Rao
authors.forename: Daniel D
authors.initials: DD
authors.lastname: Oprian
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002417:Cattle; D018698:Glutamic Acid / Q000235:genetics; D005973:Glutamine / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007532:Isoleucine / Q000235:genetics*; D007700:Kinetics; D008027:Light; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009755:Night Blindness / Q000151:congenital / Q000235:genetics* / Q000378:metabolism; D012243:Rhodopsin / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D012545:Schiff Bases / Q000737:chemistry; D013912:Threonine / Q000235:genetics*; D015293:Transducin / Q000378:metabolism; D014162:Transfection
chemical_list: D012545:Schiff Bases; D007532:Isoleucine; D005973:Glutamine; D013912:Threonine; D018698:Glutamic Acid; C013110:metarhodopsins; D012243:Rhodopsin; D015293:Transducin