title: Comparison of the phosphodiesterase inhibitory subunit interactions of frog and bovine rod outer segments.
abstract: The rod outer segments of the bovine and frog retina possess a cyclic GMP phosphodiesterase (PDE) that is composed of two larger subunits, alpha and beta (P alpha beta), which contain the catalytic activity and a smaller gamma (P gamma) subunit which inhibits the catalytic activity. We studied the binding of P gamma to P alpha beta in both the bovine and frog rod outer segment membranes. Analysis of these data indicates that there are two classes of P gamma binding sites per P alpha beta in both species. The activation of PDE by the guanosine 5'-[gamma-thio]triphosphate form of the alpha subunit of transducin, T alpha.GTP gamma S, was also studied. These data indicate that the two classes of P gamma binding sites contribute to the formation of two classes of binding sites for T alpha.GTP gamma S. We demonstrate solubilization of a portion of the P gamma by T alpha.GTP gamma S in both species. There is also present, in both species, a second class of P gamma which is not solubilized even when it is dissociated from its inhibitory site on P alpha beta by T alpha.GTP gamma S. The amount of full PDE activity which results from release of the solubilizable P gamma is about 50% in the frog PDE but only approx. 17% in the bovine PDE. We also show that activation of frog rod outer segment PDE by trypsin treatment releases the PDE from the membranes. This type of release by trypsin has already been demonstrated in bovine rod outer segments [Wensel & Stryer (1986) Proteins: Struct. Funct. Genet. 1, 90-99].
authors.affiliation: Life Sciences Division, Los Alamos National Laboratory, NM 87545.
authors.forename: M M
authors.initials: MM
authors.lastname: Whalen
authors.forename: M W
authors.initials: MW
authors.lastname: Bitensky
mesh_terms: D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002384:Catalysis; D002417:Cattle; D010786:Photoreceptor Cells / Q000201:enzymology*; D011892:Rana catesbeiana; D012374:Rod Cell Outer Segment / Q000201:enzymology*; D015293:Transducin / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
chemical_list: D015106:3',5'-Cyclic-GMP Phosphodiesterases; D014357:Trypsin; D015293:Transducin