cGMP- and phosphodiesterase-dependent light-scattering changes in rod disk membrane vesicles: relationship to disk vesicle-disk vesicle aggregation.
Visible light activates a large guanosine cyclic 3',5'-phosphate (cGMP)- and phosphodiesterase (PDE)-dependent infrared light-scattering change in suspensions of photoreceptor disk membranes. Reconstitution experiments show that this signal requires bleached rhodopsin, G protein (three polypeptide subunits of Mr 39 000, 37 000, and 6000 which comprise the GTPase), phosphodiesterase, cGMP, and GTP. The lowest light intensity which elicits the light-scattering signal bleaches 0.002% rhodopsin. cGMP and GTP hydrolysis occurs more slowly than the initial phase of the scattering signal, and the kinetics of nucleotide hydrolysis do not correlate with any phase of the signal. Hydrolysis-resistant analogues of cGMP and GTP support the initial decreasing phase of the signal. Thus, the signal apparently depends upon nucleotide binding rather than hydrolysis. Microscopic observations made under the same conditions as light-scattering experiments show that vesicle-vesicle aggregation and disaggregation occur. The data suggest that light and nucleotide activations of the cyclic nucleotide cascade enzymes are responsible for the vesicle aggregation process and nucleotide hydrolysis for vesicle disaggregation. The vesicle aggregation-disaggregation phenomenon appears likely to be the physical basis of the cGMP- and PDE-dependent changes in infrared transmission.
D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism / Q000528:radiation effects / Q000648:ultrastructure; D006152:Cyclic GMP / Q000494:pharmacology*; D005136:Eye Proteins / Q000378:metabolism*; D006150:Guanine Nucleotides / Q000494:pharmacology; D020962:Heterotrimeric GTP-Binding Proteins; D007700:Kinetics; D008027:Light; D008565:Membrane Proteins / Q000378:metabolism*; D008854:Microscopy, Electron; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D010786:Photoreceptor Cells / Q000378:metabolism / Q000528:radiation effects*; D012374:Rod Cell Outer Segment / Q000528:radiation effects*; D012542:Scattering, Radiation; D015293:Transducin
D005136:Eye Proteins; C034809:GNAT1 protein, human; D006150:Guanine Nucleotides; D008565:Membrane Proteins; D010727:Phosphoric Diester Hydrolases; D020962:Heterotrimeric GTP-Binding Proteins; D015293:Transducin; D006152:Cyclic GMP