Angiotensin II receptor recognized by DuP753 regulates two distinct guanine nucleotide-binding protein signaling pathways

Keith W. Crawford, Elizabeth A. Frey, Thomas E. Cote

Research output: Contribution to journalArticle

Abstract

The 7315c cell, derived from a rat anterior pituitary tumor, expresses an angiotensin II (AII) receptor. [3H]AII binds to 7315c membranes specifically and saturably (KD = 2.1 ± 0.6 × 10-9 M, Bmax = 282 ± 33 fmol/mg of protein). GTP diminished the affinity of the membranes for [3H]AII (KD = 4.1 ± 0.4 × 10-9 M, Bmax = 210 ± 26 fmol/mg of protein). [3H]AII binding was displaced by AII (Ki = 1.3 ± 0.6 × 10-9 M), angiotensin III (AIII) (Ki = 0.9 ± 0.4 ± 10-9 M), and the nonpeptide AII antagonist DuP753 (Ki = 1.4 ± 0.6 × 10-8 M). In contrast, a second nonpeptide AII ligand, PD123177, did not compete for [3H]AII binding sites. In intact cells, AII and AIII stimulated inositol trisphosphate (IP3) production (EC50 = 1.1 ± 0.6 × 10-8 M and 1.1 ± 0.5 × 10-8 M, respectively); this response to AII was antagonized by DuP753 (Ki = 1.7 ± 0.3 × 10-7 M). Pertussis toxin treatment failed to affect the ability of AII to stimulate IP3 production. In a crude membrane preparation, GTP was required for maximal AII-induced IP3 stimulation; guanosine thio-diphosphate abolished the agonist-GTP stimulation of IP3 production, in a concentration-dependent fashion. AII and AIII also inhibited adenylyl cyclase (EC50 = 2.9 ± 1.1 × 10-8 M and 6.0 ± 1.0 × 10-8 M, respectively). DuP753 antagonized the inhibition by AII of adenylyl cyclase (Ki = 2.8 ± 0.4 × 10-8 M). PD123177 failed to antagonize AII-induced cyclase inhibition. Pertussis toxin treatment abolished the AII and AIII inhibition of adenylyl cyclase. GTP was required for AII-induced inhibition of adenylyl cyclase. These data suggest that, in 7315c cells, a single subtype of AII receptor, identified by DuP753, is capable of regulating two different guanine nucleotide-binding protein (G protein) signalling pathways; one G protein, which is insensitive to pertussis toxin, stimulates IP3 production and the other G protein, which is sensitive to pertussis toxin, inhibits adenylyl cyclase.

Original languageEnglish (US)
Pages (from-to)154-162
Number of pages9
JournalMolecular Pharmacology
Volume41
Issue number1
StatePublished - Jan 1992
Externally publishedYes

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Angiotensin Receptors
Guanine Nucleotides
Angiotensin II
Carrier Proteins
Angiotensin III
Adenylyl Cyclases
Pertussis Toxin
Guanosine Triphosphate
GTP-Binding Proteins
Membranes
Guanosine
Diphosphates
Pituitary Neoplasms
Inositol

ASJC Scopus subject areas

  • Pharmacology

Cite this

Angiotensin II receptor recognized by DuP753 regulates two distinct guanine nucleotide-binding protein signaling pathways. / Crawford, Keith W.; Frey, Elizabeth A.; Cote, Thomas E.

In: Molecular Pharmacology, Vol. 41, No. 1, 01.1992, p. 154-162.

Research output: Contribution to journalArticle

Crawford, Keith W. ; Frey, Elizabeth A. ; Cote, Thomas E. / Angiotensin II receptor recognized by DuP753 regulates two distinct guanine nucleotide-binding protein signaling pathways. In: Molecular Pharmacology. 1992 ; Vol. 41, No. 1. pp. 154-162.
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abstract = "The 7315c cell, derived from a rat anterior pituitary tumor, expresses an angiotensin II (AII) receptor. [3H]AII binds to 7315c membranes specifically and saturably (KD = 2.1 ± 0.6 × 10-9 M, Bmax = 282 ± 33 fmol/mg of protein). GTP diminished the affinity of the membranes for [3H]AII (KD = 4.1 ± 0.4 × 10-9 M, Bmax = 210 ± 26 fmol/mg of protein). [3H]AII binding was displaced by AII (Ki = 1.3 ± 0.6 × 10-9 M), angiotensin III (AIII) (Ki = 0.9 ± 0.4 ± 10-9 M), and the nonpeptide AII antagonist DuP753 (Ki = 1.4 ± 0.6 × 10-8 M). In contrast, a second nonpeptide AII ligand, PD123177, did not compete for [3H]AII binding sites. In intact cells, AII and AIII stimulated inositol trisphosphate (IP3) production (EC50 = 1.1 ± 0.6 × 10-8 M and 1.1 ± 0.5 × 10-8 M, respectively); this response to AII was antagonized by DuP753 (Ki = 1.7 ± 0.3 × 10-7 M). Pertussis toxin treatment failed to affect the ability of AII to stimulate IP3 production. In a crude membrane preparation, GTP was required for maximal AII-induced IP3 stimulation; guanosine thio-diphosphate abolished the agonist-GTP stimulation of IP3 production, in a concentration-dependent fashion. AII and AIII also inhibited adenylyl cyclase (EC50 = 2.9 ± 1.1 × 10-8 M and 6.0 ± 1.0 × 10-8 M, respectively). DuP753 antagonized the inhibition by AII of adenylyl cyclase (Ki = 2.8 ± 0.4 × 10-8 M). PD123177 failed to antagonize AII-induced cyclase inhibition. Pertussis toxin treatment abolished the AII and AIII inhibition of adenylyl cyclase. GTP was required for AII-induced inhibition of adenylyl cyclase. These data suggest that, in 7315c cells, a single subtype of AII receptor, identified by DuP753, is capable of regulating two different guanine nucleotide-binding protein (G protein) signalling pathways; one G protein, which is insensitive to pertussis toxin, stimulates IP3 production and the other G protein, which is sensitive to pertussis toxin, inhibits adenylyl cyclase.",
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N2 - The 7315c cell, derived from a rat anterior pituitary tumor, expresses an angiotensin II (AII) receptor. [3H]AII binds to 7315c membranes specifically and saturably (KD = 2.1 ± 0.6 × 10-9 M, Bmax = 282 ± 33 fmol/mg of protein). GTP diminished the affinity of the membranes for [3H]AII (KD = 4.1 ± 0.4 × 10-9 M, Bmax = 210 ± 26 fmol/mg of protein). [3H]AII binding was displaced by AII (Ki = 1.3 ± 0.6 × 10-9 M), angiotensin III (AIII) (Ki = 0.9 ± 0.4 ± 10-9 M), and the nonpeptide AII antagonist DuP753 (Ki = 1.4 ± 0.6 × 10-8 M). In contrast, a second nonpeptide AII ligand, PD123177, did not compete for [3H]AII binding sites. In intact cells, AII and AIII stimulated inositol trisphosphate (IP3) production (EC50 = 1.1 ± 0.6 × 10-8 M and 1.1 ± 0.5 × 10-8 M, respectively); this response to AII was antagonized by DuP753 (Ki = 1.7 ± 0.3 × 10-7 M). Pertussis toxin treatment failed to affect the ability of AII to stimulate IP3 production. In a crude membrane preparation, GTP was required for maximal AII-induced IP3 stimulation; guanosine thio-diphosphate abolished the agonist-GTP stimulation of IP3 production, in a concentration-dependent fashion. AII and AIII also inhibited adenylyl cyclase (EC50 = 2.9 ± 1.1 × 10-8 M and 6.0 ± 1.0 × 10-8 M, respectively). DuP753 antagonized the inhibition by AII of adenylyl cyclase (Ki = 2.8 ± 0.4 × 10-8 M). PD123177 failed to antagonize AII-induced cyclase inhibition. Pertussis toxin treatment abolished the AII and AIII inhibition of adenylyl cyclase. GTP was required for AII-induced inhibition of adenylyl cyclase. These data suggest that, in 7315c cells, a single subtype of AII receptor, identified by DuP753, is capable of regulating two different guanine nucleotide-binding protein (G protein) signalling pathways; one G protein, which is insensitive to pertussis toxin, stimulates IP3 production and the other G protein, which is sensitive to pertussis toxin, inhibits adenylyl cyclase.

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