Assessing structural elements that influence Schiff base stability: mutants E113Q and D190N destabilize rhodopsin through different mechanisms.
The stability of the retinal chromophore attachment varies between different visual pigments and may factor in some retinal disease states. Opsin appears to stabilize this Schiff base linkage by: (i) affecting the hydrolysis chemistry, (ii) shielding the retinal linkage from solvent, or (iii) acting as a kinetic trap to slow retinal release. Here we describe methods to determine Schiff base stability in rhodopsin, present examples of dark state and MII rhodopsin stability differences, and show that studies of mutants E113Q and D190N demonstrate different parts of rhodopsin influence Schiff base stability in different ways.
Janz
Jay M
JM
Department of Biochemistry and Molecular Biology, Oregon Health and Science University, Mail Code L224 3181 S.W. Sam Jackson Park Road, Portland, OR 97239, USA.
Farrens
David L
DL
D008040:Genetic Linkage; D006801:Humans; D006868:Hydrolysis; D019811:Hydroxylamine / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D012160:Retina / Q000378:metabolism*; D012243:Rhodopsin / Q000235:genetics* / Q000378:metabolism; D012545:Schiff Bases / Q000737:chemistry*; D013057:Spectrum Analysis / Q000379:methods
D012545:Schiff Bases; D019811:Hydroxylamine; D012243:Rhodopsin