Identification of surrogate ligands for orphan G protein-coupled receptors.
We prepared fusion proteins with an alpha subunit of G protein Gi (Gi1alpha) of 26 orphan G protein-coupled receptors (GPCRs) and with Gsalpha of 10 orphan GPCRs, most of which had been identified from the human genome previously [FEBS Lett 520 (2002) 97]. Ligands for these fusion proteins were screened from a library consisting of approximately 1000 authentic compounds by measuring their effect on [35S]GTPgammaS binding to membrane preparations of insect Sf9 cells expressing these fusion proteins. Eleven compounds were found to act as surrogate agonists for a GPCR-Gsalpha and four GPCR-Gialpha fusion proteins, a compound as an inverse agonist for two GPCR-Gsalpha fusion proteins, and a compound as an endogenous agonist for a GPCR-Gialpha fusion protein.
Takeda
Shigeki
S
Institute for Biomolecular Science, Gakushuin University, 1-5-1 Mejiro, Tokyo 171-8588, Toshima-ku, Japan. tasuya.haga@gakushuin.ac.jp
Yamamoto
Atsushi
A
Okada
Tomoaki
T
Matsumura
Emiko
E
Nose
Emi
E
Kogure
Katsuya
K
Kojima
Shigen
S
Haga
Tatsuya
T
D000818:Animals; D015153:Blotting, Western; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000235:genetics / Q000502:physiology; D015723:Gene Library; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006153:Guanosine Diphosphate / Q000494:pharmacology; D006801:Humans; D007313:Insecta; D008024:Ligands; D043562:Receptors, G-Protein-Coupled / Q000096:biosynthesis / Q000187:drug effects* / Q000235:genetics; D011993:Recombinant Fusion Proteins
D018076:DNA, Complementary; D008024:Ligands; D043562:Receptors, G-Protein-Coupled; D011993:Recombinant Fusion Proteins; D006153:Guanosine Diphosphate; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D019205:GTP-Binding Protein alpha Subunits, Gs