Light- and GTP-activated hydrolysis of phosphatidylinositol bisphosphate in squid photoreceptor membranes.
Light stimulates the hydrolysis of exogenous, [3H]inositol-labeled phosphatidylinositol bisphosphate (PtdInsP2) added to squid photoreceptor membranes, releasing inositol trisphosphate (InsP3). At free calcium levels of 0.05 microM or greater, hydrolysis of the labeled lipid is stimulated up to 4-fold by GTP and light together, but not separately. This activity is the biochemical counterpart of observations on intact retina showing that a rhodopsin-activated GTP-binding protein is involved in visual transduction in invertebrates, and that InsP3 release is correlated with visual excitation and adaptation. Using an in vitro assay, we investigated the calcium and GTP dependence of the phospholipase activity. At calcium concentrations between 0.1 and 0.5 microM, some hydrolysis occurs independently of GTP and light, with a light- and GTP-activated component superimposed. At 1 microM calcium there is no background activity, and hydrolysis absolutely requires both GTP and light. Ion exchange chromatography on Dowex 1 (formate form) of the water-soluble products released at 1 microM calcium reveals that the product is almost entirely InsP3. Invertebrate rhodopsin is homologous in sequence and function to vertebrate visual pigment, which modulates the concentration of cyclic GMP through the mediation of the GTP-binding protein transducin. While there is some evidence that light also modulates PtdInsP2 content in vertebrate photoreceptors, the case for its involvement in phototransduction is stronger for the invertebrate systems. The results reported here support the scheme of rhodopsin----GTP-binding protein----phospholipase C activation in invertebrate photoreceptors.
Baer
K M
KM
Department of Zoology, University of Oxford, United Kingdom.
Saibil
H R
HR
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism / Q000528:radiation effects; D049832:Decapodiformes; D006160:Guanosine Triphosphate / Q000494:pharmacology*; D007700:Kinetics; D008027:Light; D019269:Phosphatidylinositol 4,5-Diphosphate; D010716:Phosphatidylinositols / Q000378:metabolism* / Q000528:radiation effects; D010786:Photoreceptor Cells / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects
D019269:Phosphatidylinositol 4,5-Diphosphate; D010716:Phosphatidylinositols; D006160:Guanosine Triphosphate