Stimulatory effect of guanine nucleotides on prostaglandin E1 binding to murine renal outer medulla.
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.
Kimura
K
K
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Fujiwara
N
N
Negishi
M
M
Yatsunami
K
K
Ichikawa
A
A
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
D003432:Cross-Linking Reagents; D006150:Guanine Nucleotides; D011982:Receptors, Prostaglandin; D000527:Alprostadil