Unit

UNIT 2.2 Adenylyl and Guanylyl Cyclase Assays

  1. John J. Kelly1,
  2. Troy Stevens1,
  3. W. Joseph Thompson2,
  4. Roland Seifert3

Published Online: 1 OCT 2005

DOI: 10.1002/0471141755.ph0202s30

Current Protocols in Pharmacology

Current Protocols in Pharmacology

How to Cite

Kelly, J. J., Stevens, T., Thompson, W. J. and Seifert, R. 2005. Adenylyl and Guanylyl Cyclase Assays. Current Protocols in Pharmacology. 30:2.2:2.2.1–2.2.21.

Author Information

  1. 1

    University of South Alabama College of Medicine, Mobile, Alabama

  2. 2

    Cell Pathways, Horsham, Pennsylvania

  3. 3

    Universität Regensburg, Regensburg, Germany

Publication History

  1. Published Online: 1 OCT 2005
  2. Published Print: SEP 2005

Abstract

This unit presents two basic protocols to determine adenylyl cyclase and guanylyl cyclase activity in tissue and cell homogenates, permeabilized cells, or subcellular fractions. Each method is divided into two parts: the enzyme reaction that causes the formation of the labeled cyclic nucleotide, and the separation of cyclic nucleotide products from unreacted nucleotide triphosphates and metabolites using Dowex 50 resin and aluminum oxide chromatographies. In the case of guanylyl cyclase, alternative separation protocols are also provided. Additionally, protocols are provided that describe preparation of both the columns used in the assays and the tissue or cells to be assayed.

Keywords:

  • cyclase;
  • cAMP;
  • cGMP;
  • G proteins;
  • G protein-coupled receptors;
  • cyclic nucleotide