Adenylate cyclase in the corpus luteum of the rhesus monkey. I. General properties and optimal assay conditions

K. M. Eyster, Richard Stouffer

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

To characterize the adenylate cyclase system of the primate corpus luteum, we assayed the conversion of [α-32P] ATP to [32P]cAMP in preparations of luteal tissue obtained from rhesus monkeys on days 17-19 of the menstrual cycle. Basal, gonadotropin (hCG; 250 nM)- sensitive, and guanine nucleotide (5'-guanylyl-imidodiphosphate [GMP-P(NH)P]; 10 μM)-sensitive cAMP production were influenced by the pH, osmolality, and ionic strength of the assay buffer. As the concentration of Mg2+ increased from 2 to 10 mM, adenylate cyclase activity was enhanced; however, the relative stimulation by hCG plus GMP-P(NH)P was maximal when the Mg2+ concentration approximated that of ATP plus EDTA. In contrast, the presence of Ca+2 inhibited basal, hCG-stimulated, and GMP-P(NH)P-stimulated cAMP production. Adenylate cyclase activity was substrate dependent at ATP concentrations from 0.7-4.5 mM; however, higher concentrations of ATP did not alter cAMP production. The relative stimulation by hCG and GMP-P(NH)P was independent of ATP levels when the ATP to Mg ratio was constant. The rate of cAMP production was constant during 30 min of incubation at 37 C, with the ATP concentration maintained at greater than 87% of initial levels. Adenylate cyclase activity was 10-fold greater in luteal tissue from the superovulated rat than in that from the cycling rhesus monkey; however, relative stimulation by hCG and GMP-P(NH)P was qualitatively similar in the two species. Thus, we have demonstrated the existence of an adenylate cyclase system in the corpus luteum of the rhesus monkey during the menstrual cycle and have established some general properties and optimal assay conditions for the gonadotropin-sensitive adenylate cyclase.

Original languageEnglish (US)
Pages (from-to)1543-1551
Number of pages9
JournalEndocrinology
Volume116
Issue number4
StatePublished - 1985
Externally publishedYes

Fingerprint

Guanylyl Imidodiphosphate
Corpus Luteum
Macaca mulatta
Adenylyl Cyclases
Adenosine Triphosphate
Menstrual Cycle
Gonadotropins
Osmolar Concentration
Guanine Nucleotides
Edetic Acid
Primates
Buffers

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Adenylate cyclase in the corpus luteum of the rhesus monkey. I. General properties and optimal assay conditions. / Eyster, K. M.; Stouffer, Richard.

In: Endocrinology, Vol. 116, No. 4, 1985, p. 1543-1551.

Research output: Contribution to journalArticle

@article{1b3cdff1f0d84717bb36620a33d49eea,
title = "Adenylate cyclase in the corpus luteum of the rhesus monkey. I. General properties and optimal assay conditions",
abstract = "To characterize the adenylate cyclase system of the primate corpus luteum, we assayed the conversion of [α-32P] ATP to [32P]cAMP in preparations of luteal tissue obtained from rhesus monkeys on days 17-19 of the menstrual cycle. Basal, gonadotropin (hCG; 250 nM)- sensitive, and guanine nucleotide (5'-guanylyl-imidodiphosphate [GMP-P(NH)P]; 10 μM)-sensitive cAMP production were influenced by the pH, osmolality, and ionic strength of the assay buffer. As the concentration of Mg2+ increased from 2 to 10 mM, adenylate cyclase activity was enhanced; however, the relative stimulation by hCG plus GMP-P(NH)P was maximal when the Mg2+ concentration approximated that of ATP plus EDTA. In contrast, the presence of Ca+2 inhibited basal, hCG-stimulated, and GMP-P(NH)P-stimulated cAMP production. Adenylate cyclase activity was substrate dependent at ATP concentrations from 0.7-4.5 mM; however, higher concentrations of ATP did not alter cAMP production. The relative stimulation by hCG and GMP-P(NH)P was independent of ATP levels when the ATP to Mg ratio was constant. The rate of cAMP production was constant during 30 min of incubation at 37 C, with the ATP concentration maintained at greater than 87{\%} of initial levels. Adenylate cyclase activity was 10-fold greater in luteal tissue from the superovulated rat than in that from the cycling rhesus monkey; however, relative stimulation by hCG and GMP-P(NH)P was qualitatively similar in the two species. Thus, we have demonstrated the existence of an adenylate cyclase system in the corpus luteum of the rhesus monkey during the menstrual cycle and have established some general properties and optimal assay conditions for the gonadotropin-sensitive adenylate cyclase.",
author = "Eyster, {K. M.} and Richard Stouffer",
year = "1985",
language = "English (US)",
volume = "116",
pages = "1543--1551",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "4",

}

TY - JOUR

T1 - Adenylate cyclase in the corpus luteum of the rhesus monkey. I. General properties and optimal assay conditions

AU - Eyster, K. M.

AU - Stouffer, Richard

PY - 1985

Y1 - 1985

N2 - To characterize the adenylate cyclase system of the primate corpus luteum, we assayed the conversion of [α-32P] ATP to [32P]cAMP in preparations of luteal tissue obtained from rhesus monkeys on days 17-19 of the menstrual cycle. Basal, gonadotropin (hCG; 250 nM)- sensitive, and guanine nucleotide (5'-guanylyl-imidodiphosphate [GMP-P(NH)P]; 10 μM)-sensitive cAMP production were influenced by the pH, osmolality, and ionic strength of the assay buffer. As the concentration of Mg2+ increased from 2 to 10 mM, adenylate cyclase activity was enhanced; however, the relative stimulation by hCG plus GMP-P(NH)P was maximal when the Mg2+ concentration approximated that of ATP plus EDTA. In contrast, the presence of Ca+2 inhibited basal, hCG-stimulated, and GMP-P(NH)P-stimulated cAMP production. Adenylate cyclase activity was substrate dependent at ATP concentrations from 0.7-4.5 mM; however, higher concentrations of ATP did not alter cAMP production. The relative stimulation by hCG and GMP-P(NH)P was independent of ATP levels when the ATP to Mg ratio was constant. The rate of cAMP production was constant during 30 min of incubation at 37 C, with the ATP concentration maintained at greater than 87% of initial levels. Adenylate cyclase activity was 10-fold greater in luteal tissue from the superovulated rat than in that from the cycling rhesus monkey; however, relative stimulation by hCG and GMP-P(NH)P was qualitatively similar in the two species. Thus, we have demonstrated the existence of an adenylate cyclase system in the corpus luteum of the rhesus monkey during the menstrual cycle and have established some general properties and optimal assay conditions for the gonadotropin-sensitive adenylate cyclase.

AB - To characterize the adenylate cyclase system of the primate corpus luteum, we assayed the conversion of [α-32P] ATP to [32P]cAMP in preparations of luteal tissue obtained from rhesus monkeys on days 17-19 of the menstrual cycle. Basal, gonadotropin (hCG; 250 nM)- sensitive, and guanine nucleotide (5'-guanylyl-imidodiphosphate [GMP-P(NH)P]; 10 μM)-sensitive cAMP production were influenced by the pH, osmolality, and ionic strength of the assay buffer. As the concentration of Mg2+ increased from 2 to 10 mM, adenylate cyclase activity was enhanced; however, the relative stimulation by hCG plus GMP-P(NH)P was maximal when the Mg2+ concentration approximated that of ATP plus EDTA. In contrast, the presence of Ca+2 inhibited basal, hCG-stimulated, and GMP-P(NH)P-stimulated cAMP production. Adenylate cyclase activity was substrate dependent at ATP concentrations from 0.7-4.5 mM; however, higher concentrations of ATP did not alter cAMP production. The relative stimulation by hCG and GMP-P(NH)P was independent of ATP levels when the ATP to Mg ratio was constant. The rate of cAMP production was constant during 30 min of incubation at 37 C, with the ATP concentration maintained at greater than 87% of initial levels. Adenylate cyclase activity was 10-fold greater in luteal tissue from the superovulated rat than in that from the cycling rhesus monkey; however, relative stimulation by hCG and GMP-P(NH)P was qualitatively similar in the two species. Thus, we have demonstrated the existence of an adenylate cyclase system in the corpus luteum of the rhesus monkey during the menstrual cycle and have established some general properties and optimal assay conditions for the gonadotropin-sensitive adenylate cyclase.

UR - http://www.scopus.com/inward/record.url?scp=0021799923&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021799923&partnerID=8YFLogxK

M3 - Article

C2 - 3971926

AN - SCOPUS:0021799923

VL - 116

SP - 1543

EP - 1551

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 4

ER -