Chapter 8. Methodological Approaches

  1. Prof. Roland Seifert1 and
  2. Prof. Thomas Wieland2
  1. Prof. Thomas Wieland2 and
  2. Prof. Roland Seifert1

Published Online: 25 APR 2006

DOI: 10.1002/352760734X.ch8

G Protein-Coupled Receptors as Drug Targets: Analysis of Activation and Constitutive Activity, Volume 24

G Protein-Coupled Receptors as Drug Targets: Analysis of Activation and Constitutive Activity, Volume 24

How to Cite

Wieland, T. and Seifert, R. (2005) Methodological Approaches, in G Protein-Coupled Receptors as Drug Targets: Analysis of Activation and Constitutive Activity, Volume 24 (eds R. Seifert and T. Wieland), Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, FRG. doi: 10.1002/352760734X.ch8

Editor Information

  1. 1

    Department of Pharmacology and Toxicology, University of Regensburg, Universitätsstraße 31, D-93053 Regensburg, Germany

  2. 2

    Institut für Pharmakologie und Toxikologie, Fakultät für Klinische Medizin Mannheim der Universität Heidelberg, Maybachstraße 14-16, 68169 Mannheim, Germany

Author Information

  1. 1

    Department of Pharmacology and Toxicology, University of Regensburg, Universitätsstraße 31, D-93053 Regensburg, Germany

  2. 2

    Institut für Pharmakologie und Toxikologie, Fakultät für Klinische Medizin Mannheim der Universität Heidelberg, Maybachstraße 14-16, 68169 Mannheim, Germany

Publication History

  1. Published Online: 25 APR 2006
  2. Published Print: 26 SEP 2005

Book Series:

  1. Methods and Principles in Medicinal Chemistry

Book Series Editors:

  1. Prof. Dr. Raimund Mannhold3,
  2. Prof. Dr. Hugo Kubinyi4 and
  3. Prof. Dr. Gerd Folkers5

Series Editor Information

  1. 3

    Biomedical Research Center, Molecular Drug Research Group, Heinrich-Heine-Universität, Universitätsstrasse 1, 40225 Düsseldorf, Germany

  2. 4

    Donnersbergstrasse 9, 67256 Weisenheim am Sand, Germany

  3. 5

    Collegium Helveticum, STW/ETH Zentrum, 8092 Zürich, Switzerland

ISBN Information

Print ISBN: 9783527308194

Online ISBN: 9783527607341

SEARCH

Keywords:

  • G protein coupled receptors (GPCR);
  • methodological approaches;
  • measurement of constitutive GPCR activity;
  • GPCR activity in membranes;
  • GPCR activity intact cells;
  • determination of cAMP concentrations;
  • determination of inositol phosphate formation;
  • SRF-mediated gene transcription;
  • deorphanization;
  • aequorin-based Ca2+ determinations

Summary

This chapter contains sections titled:

  • Introduction

  • Analysis of Constitutive GPCR Activity in Membranes and Intact Cells

    • Procedure for Sf9 Cell Culture and Membrane Preparation

    • GPCR Radioligand Binding Studies

    • GTPase Assay

    • [35S]GTPγS Binding Assay

    • Adenylyl Cyclase Assay

  • Measurement of Constitutive Activity of GPCRs in Intact Cells

    • Quantitative Determination of cAMP Concentrations in Cell Culture Lysates

    • Determination of Inositol Phosphate Formation in Living Cells

    • Determination of G Protein Activation by SRF-mediated Gene Transcription

    • Deorphanization and Constitutive Activity of GPCRs by Aequorin-based Ca2+ Determinations