Light-regulated biochemical events in invertebrate photoreceptors. 1. Light-activated guanosinetriphosphatase, guanine nucleotide binding, and cholera toxin catalyzed labeling of squid photoreceptor membranes.
The occurrence of a guanine nucleotide binding protein activated by squid rhodopsin was established by examination of GTPase activity, guanine nucleotide binding, and cholera toxin catalyzed labeling of squid photoreceptor membranes. Purified squid (Loligo opalescens) photoreceptors exhibited GTPase activity that increased 3-4-fold by illumination. Half-maximal GTPase activity was observed when 2% of the rhodopsin was photoconverted to metarhodopsin. The Km of the light-regulated activity was 1 microM GTP. Binding of the hydrolysis-resistant GTP analogue guanosine 5'-(beta, gamma-imidotriphosphate) [Gpp(NH)p] was enhanced greater than 10 times by illumination. A protein, Mr 44 000, was identified as a component of the light-activated guanine nucleotide binding protein/GTPase through its specific labeling with [32P]NAD catalyzed by cholera toxin: light increased the extent of 32P incorporation 7-fold. The addition of ATP to the membrane suspension enhanced labeling, while guanine nucleotides inhibited labeling with the relative potency GTP gamma S much greater than GDP greater than GTP greater than Gpp(NH)p. The 44 000-dalton protein was membrane bound irrespective of variations in ionic strength and divalent ion concentration over a wide range. These results suggest that a G protein, which incorporates both GTP binding and hydrolysis functions, is intimately involved in the visual process of invertebrate photoreceptors.
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002772:Cholera Toxin / Q000494:pharmacology*; D049832:Decapodiformes; D020558:GTP Phosphohydrolases / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006150:Guanine Nucleotides / Q000494:pharmacology; D007700:Kinetics; D008027:Light; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D010786:Photoreceptor Cells / Q000187:drug effects / Q000378:metabolism*
D002352:Carrier Proteins; D006150:Guanine Nucleotides; D002772:Cholera Toxin; D010744:Phosphoric Monoester Hydrolases; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins