title: Stimulatory effect of guanine nucleotides on prostaglandin E1 binding to murine renal outer medulla.
abstract: Prostaglandin E1 (PGE1) specifically bound to the membrane prepared from murine renal outer medulla. The extent of binding of [3H]PGE1 to the membrane was increased about 4-fold by guanosine triphosphate (GTP) and its analogs, but the dissociation of bound [3H]PGE1 from the membrane was in turn enhanced by GTP gamma S. Scatchard plot analyses revealed that GTP gamma S increased the binding affinity more than 2-fold without a major change in the number of binding sites. When [3H]PGE1-bound proteins were cross-linked in the membrane by dithiobis (succinimidyl propionate), bound [3H]PGE1 was no longer dissociated by GTP gamma S treatment, suggesting that cross-linking produced a stable complex of PGE receptor with a GTP-binding protein. The cross-linked [3H]PGE1-specifically bound proteins solubilized from the membranes labeled with [3H] PGE1 in the presence or absence of GTP gamma S were eluted as an apparently single radioactive peak at the same position of Mr = 15000 by gel filtration, indicating that the PGE receptor forms a complex with a GTP-binding protein regardless of the treatment with GTP gamma S by which [3H]PGE1 binding is promoted. The ability of GTP gamma S to stimulate [3H]PGE1 binding was eliminated by pretreatment of the membrane with pertussis toxin, but not cholera toxin, indicating that the PGE receptor is coupled to a pertussis toxin-sensitive GTP-binding protein.
authors.affiliation: Department of Biochemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Kimura
authors.forename: N
authors.initials: N
authors.lastname: Fujiwara
authors.forename: M
authors.initials: M
authors.lastname: Negishi
authors.forename: K
authors.initials: K
authors.lastname: Yatsunami
authors.forename: A
authors.initials: A
authors.lastname: Ichikawa
mesh_terms: D000527:Alprostadil / Q000378:metabolism*; D000818:Animals; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents; D006150:Guanine Nucleotides / Q000494:pharmacology*; D066298:In Vitro Techniques; D007679:Kidney Medulla / Q000378:metabolism*; D008566:Membranes / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D011982:Receptors, Prostaglandin / Q000378:metabolism*; D012995:Solubility
chemical_list: D003432:Cross-Linking Reagents; D006150:Guanine Nucleotides; D011982:Receptors, Prostaglandin; D000527:Alprostadil