A comparison of the efficiency of G protein activation by ligand-free and light-activated forms of rhodopsin.
Activation of the photoreceptor G protein transducin (Gt) by opsin, the ligand-free form of rhodopsin, was measured using rod outer segment membranes with densities of opsin and Gt similar to those found in rod cells. When GTPgammaS was used as the activating nucleotide, opsin catalyzed transducin activation with an exponential time course with a rate constant k(act) on the order of 2 x 10(-3)s(-1). Comparison under these conditions to activation by flash-generated metarhodopsin II (MII) revealed that opsin- and R*-catalyzed activation showed similar kinetics when MII was present at a surface density approximately 10(-6) lower than that of opsin. Thus, in contrast to some previous reports, we find that the catalytic potency of opsin is only approximately 10(-6) that of MII. In the presence of residual retinaldehyde-derived species present in membranes treated with hydroxylamine after bleaching, the apparent k(act) observed was much higher than that for opsin, suggesting a possible explanation for previous reports of more efficient activation by opsin. These results are important for considering the possible role of opsin in the diverse phenomena in which it has been suggested to play a key role, such as bleaching desensitization and retinal degeneration induced by continuous light or vitamin A deprivation.
Melia
T J
TJ
Department of Biochemistry, Baylor College of Medicine, Houston, Texas 77030, USA.
Cowan
C W
CW
Angleson
J K
JK
Wensel
T G
TG
D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D002417:Cattle; D004224:Diterpenes; D019204:GTP-Binding Proteins / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000494:pharmacology; D066298:In Vitro Techniques; D008024:Ligands; D008027:Light; D010091:Oximes / Q000378:metabolism; D010777:Photochemistry; D012172:Retinaldehyde / Q000378:metabolism; D012243:Rhodopsin / Q000737:chemistry / Q000378:metabolism* / Q000528:radiation effects*; D012374:Rod Cell Outer Segment / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D017299:Rod Opsins / Q000378:metabolism; D015293:Transducin / Q000378:metabolism
D004224:Diterpenes; D008024:Ligands; D010091:Oximes; D017299:Rod Opsins; C030628:retinaloxime; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D012243:Rhodopsin; D019204:GTP-Binding Proteins; D015293:Transducin; D012172:Retinaldehyde