Identification of a G(iα) binding site on type V adenylyl cyclase

Carmen W. Dessauer, John J G Tesmer, Stephen R. Sprang, Alfred G. Gilman

Research output: Contribution to journalArticle

107 Citations (Scopus)

Abstract

The stimulatory G protein α subunit G(sα) binds within a cleft in adenylyl cyclase formed by the α1-α2 and α3-β4 loops of the C2 domain. The pseudosymmetry of the C1 and C2 domains of adenylyl cyclase suggests that the homologous inhibitory α subunit G(iα) could bind to the analogous cleft within C1. We demonstrate that myristoylated guanosine 5'-3-O- (thio)triphosphate-G(iα1) forms a stable complex with the C1 (but not the C2) domain of type V adenylyl cyclase. Mutagenesis of the membrane-bound enzyme identified residues whose alteration either increased or substantially decreased the IC50 for inhibition by G(iα1). These mutations suggest binding of G(iα) within the cleft formed by the α2 and α3 helices of C1, analogous to the G(sα) binding site in C2. Adenylyl cyclase activity reconstituted by mixture of the C1 and C2 domains of type V adenylyl cyclase was also inhibited by G(iα). The C(1b) domain of the type V enzyme contributed to affinity for G(iα), but the source of C2 had little effect. Mutations in this soluble system faithfully reflected the phenotypes observed with the membrane-bound enzyme. The pseudosymmetrical structure of adenylyl cyclase permits bidirectional regulation of activity by homologous G protein α subunits.

Original languageEnglish (US)
Pages (from-to)25831-25839
Number of pages9
JournalJournal of Biological Chemistry
Volume273
Issue number40
DOIs
StatePublished - Oct 2 1998

Fingerprint

Adenylyl Cyclases
Binding Sites
Protein Subunits
GTP-Binding Proteins
Enzymes
Guanosine 5'-O-(3-Thiotriphosphate)
Membranes
Mutagenesis
Mutation
Inhibitory Concentration 50
Phenotype
adenylyl cyclase type V
C2 Domains

ASJC Scopus subject areas

  • Biochemistry

Cite this

Dessauer, C. W., Tesmer, J. J. G., Sprang, S. R., & Gilman, A. G. (1998). Identification of a G(iα) binding site on type V adenylyl cyclase. Journal of Biological Chemistry, 273(40), 25831-25839. https://doi.org/10.1074/jbc.273.40.25831

Identification of a G(iα) binding site on type V adenylyl cyclase. / Dessauer, Carmen W.; Tesmer, John J G; Sprang, Stephen R.; Gilman, Alfred G.

In: Journal of Biological Chemistry, Vol. 273, No. 40, 02.10.1998, p. 25831-25839.

Research output: Contribution to journalArticle

Dessauer, CW, Tesmer, JJG, Sprang, SR & Gilman, AG 1998, 'Identification of a G(iα) binding site on type V adenylyl cyclase', Journal of Biological Chemistry, vol. 273, no. 40, pp. 25831-25839. https://doi.org/10.1074/jbc.273.40.25831
Dessauer, Carmen W. ; Tesmer, John J G ; Sprang, Stephen R. ; Gilman, Alfred G. / Identification of a G(iα) binding site on type V adenylyl cyclase. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 40. pp. 25831-25839.
@article{b266f0029a6e4c2ba12d297f2e5563d1,
title = "Identification of a G(iα) binding site on type V adenylyl cyclase",
abstract = "The stimulatory G protein α subunit G(sα) binds within a cleft in adenylyl cyclase formed by the α1-α2 and α3-β4 loops of the C2 domain. The pseudosymmetry of the C1 and C2 domains of adenylyl cyclase suggests that the homologous inhibitory α subunit G(iα) could bind to the analogous cleft within C1. We demonstrate that myristoylated guanosine 5'-3-O- (thio)triphosphate-G(iα1) forms a stable complex with the C1 (but not the C2) domain of type V adenylyl cyclase. Mutagenesis of the membrane-bound enzyme identified residues whose alteration either increased or substantially decreased the IC50 for inhibition by G(iα1). These mutations suggest binding of G(iα) within the cleft formed by the α2 and α3 helices of C1, analogous to the G(sα) binding site in C2. Adenylyl cyclase activity reconstituted by mixture of the C1 and C2 domains of type V adenylyl cyclase was also inhibited by G(iα). The C(1b) domain of the type V enzyme contributed to affinity for G(iα), but the source of C2 had little effect. Mutations in this soluble system faithfully reflected the phenotypes observed with the membrane-bound enzyme. The pseudosymmetrical structure of adenylyl cyclase permits bidirectional regulation of activity by homologous G protein α subunits.",
author = "Dessauer, {Carmen W.} and Tesmer, {John J G} and Sprang, {Stephen R.} and Gilman, {Alfred G.}",
year = "1998",
month = "10",
day = "2",
doi = "10.1074/jbc.273.40.25831",
language = "English (US)",
volume = "273",
pages = "25831--25839",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "40",

}

TY - JOUR

T1 - Identification of a G(iα) binding site on type V adenylyl cyclase

AU - Dessauer, Carmen W.

AU - Tesmer, John J G

AU - Sprang, Stephen R.

AU - Gilman, Alfred G.

PY - 1998/10/2

Y1 - 1998/10/2

N2 - The stimulatory G protein α subunit G(sα) binds within a cleft in adenylyl cyclase formed by the α1-α2 and α3-β4 loops of the C2 domain. The pseudosymmetry of the C1 and C2 domains of adenylyl cyclase suggests that the homologous inhibitory α subunit G(iα) could bind to the analogous cleft within C1. We demonstrate that myristoylated guanosine 5'-3-O- (thio)triphosphate-G(iα1) forms a stable complex with the C1 (but not the C2) domain of type V adenylyl cyclase. Mutagenesis of the membrane-bound enzyme identified residues whose alteration either increased or substantially decreased the IC50 for inhibition by G(iα1). These mutations suggest binding of G(iα) within the cleft formed by the α2 and α3 helices of C1, analogous to the G(sα) binding site in C2. Adenylyl cyclase activity reconstituted by mixture of the C1 and C2 domains of type V adenylyl cyclase was also inhibited by G(iα). The C(1b) domain of the type V enzyme contributed to affinity for G(iα), but the source of C2 had little effect. Mutations in this soluble system faithfully reflected the phenotypes observed with the membrane-bound enzyme. The pseudosymmetrical structure of adenylyl cyclase permits bidirectional regulation of activity by homologous G protein α subunits.

AB - The stimulatory G protein α subunit G(sα) binds within a cleft in adenylyl cyclase formed by the α1-α2 and α3-β4 loops of the C2 domain. The pseudosymmetry of the C1 and C2 domains of adenylyl cyclase suggests that the homologous inhibitory α subunit G(iα) could bind to the analogous cleft within C1. We demonstrate that myristoylated guanosine 5'-3-O- (thio)triphosphate-G(iα1) forms a stable complex with the C1 (but not the C2) domain of type V adenylyl cyclase. Mutagenesis of the membrane-bound enzyme identified residues whose alteration either increased or substantially decreased the IC50 for inhibition by G(iα1). These mutations suggest binding of G(iα) within the cleft formed by the α2 and α3 helices of C1, analogous to the G(sα) binding site in C2. Adenylyl cyclase activity reconstituted by mixture of the C1 and C2 domains of type V adenylyl cyclase was also inhibited by G(iα). The C(1b) domain of the type V enzyme contributed to affinity for G(iα), but the source of C2 had little effect. Mutations in this soluble system faithfully reflected the phenotypes observed with the membrane-bound enzyme. The pseudosymmetrical structure of adenylyl cyclase permits bidirectional regulation of activity by homologous G protein α subunits.

UR - http://www.scopus.com/inward/record.url?scp=0032475516&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032475516&partnerID=8YFLogxK

U2 - 10.1074/jbc.273.40.25831

DO - 10.1074/jbc.273.40.25831

M3 - Article

C2 - 9748257

AN - SCOPUS:0032475516

VL - 273

SP - 25831

EP - 25839

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 40

ER -