Adenylate cyclase assay with adenylyl imidodiphosphate and product detection by competitive protein binding

Michael E. Maguire, Alfred G. Gilman

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

An assay for adenylate cyclase (EC 4.6.1.1) is described using adenylyl imidodiphosphate (AMP-PNP) as substrate. Methods are described for introducing the cyclase reaction mixture directly into a protein binding assay for the cyclic nucleotide without purification of cyclic AMP. Significant substrate depletion and regenerating systems are avoided by this method, and blank values are negligible. The assay is capable of reproducibly detecting adenylate cyclase activity with less than 5 μg of protein from rat cerebral cortex.

Original languageEnglish (US)
Pages (from-to)154-163
Number of pages10
JournalBBA - Enzymology
Volume358
Issue number1
DOIs
StatePublished - Jul 17 1974

Fingerprint

Adenylyl Imidodiphosphate
Competitive Binding
Adenylyl Cyclases
Protein Binding
Cyclic Nucleotides
Cerebral Cortex
Cyclic AMP
Proteins

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Adenylate cyclase assay with adenylyl imidodiphosphate and product detection by competitive protein binding. / Maguire, Michael E.; Gilman, Alfred G.

In: BBA - Enzymology, Vol. 358, No. 1, 17.07.1974, p. 154-163.

Research output: Contribution to journalArticle

Maguire, Michael E. ; Gilman, Alfred G. / Adenylate cyclase assay with adenylyl imidodiphosphate and product detection by competitive protein binding. In: BBA - Enzymology. 1974 ; Vol. 358, No. 1. pp. 154-163.
@article{b38e0682ea27464b8df9c788d08d1fb4,
title = "Adenylate cyclase assay with adenylyl imidodiphosphate and product detection by competitive protein binding",
abstract = "An assay for adenylate cyclase (EC 4.6.1.1) is described using adenylyl imidodiphosphate (AMP-PNP) as substrate. Methods are described for introducing the cyclase reaction mixture directly into a protein binding assay for the cyclic nucleotide without purification of cyclic AMP. Significant substrate depletion and regenerating systems are avoided by this method, and blank values are negligible. The assay is capable of reproducibly detecting adenylate cyclase activity with less than 5 μg of protein from rat cerebral cortex.",
author = "Maguire, {Michael E.} and Gilman, {Alfred G.}",
year = "1974",
month = "7",
day = "17",
doi = "10.1016/0005-2744(74)90267-8",
language = "English (US)",
volume = "358",
pages = "154--163",
journal = "BBA - Enzymology",
issn = "0005-2744",
publisher = "Elsevier BV",
number = "1",

}

TY - JOUR

T1 - Adenylate cyclase assay with adenylyl imidodiphosphate and product detection by competitive protein binding

AU - Maguire, Michael E.

AU - Gilman, Alfred G.

PY - 1974/7/17

Y1 - 1974/7/17

N2 - An assay for adenylate cyclase (EC 4.6.1.1) is described using adenylyl imidodiphosphate (AMP-PNP) as substrate. Methods are described for introducing the cyclase reaction mixture directly into a protein binding assay for the cyclic nucleotide without purification of cyclic AMP. Significant substrate depletion and regenerating systems are avoided by this method, and blank values are negligible. The assay is capable of reproducibly detecting adenylate cyclase activity with less than 5 μg of protein from rat cerebral cortex.

AB - An assay for adenylate cyclase (EC 4.6.1.1) is described using adenylyl imidodiphosphate (AMP-PNP) as substrate. Methods are described for introducing the cyclase reaction mixture directly into a protein binding assay for the cyclic nucleotide without purification of cyclic AMP. Significant substrate depletion and regenerating systems are avoided by this method, and blank values are negligible. The assay is capable of reproducibly detecting adenylate cyclase activity with less than 5 μg of protein from rat cerebral cortex.

UR - http://www.scopus.com/inward/record.url?scp=0016169228&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0016169228&partnerID=8YFLogxK

U2 - 10.1016/0005-2744(74)90267-8

DO - 10.1016/0005-2744(74)90267-8

M3 - Article

C2 - 4368377

AN - SCOPUS:0016169228

VL - 358

SP - 154

EP - 163

JO - BBA - Enzymology

JF - BBA - Enzymology

SN - 0005-2744

IS - 1

ER -