Signal Transduction: A Practical Approach

Paperback | August 1, 1999

EditorGraeme Milligan

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Since the publication of the first edition of Signal Transduction: A Practical Approach in 1992 there has been a great deal of new information about the processes of signal transduction and consequently many new methods have been developed. This new edition has therefore been updated andextended to include the major new methods now available. The first part of the book is mainly concerned with G protein-coupled receptors and covers structural studies of conformational changes and binding sites, phosphorylation and desensitisation, identification, receptor fusion proteins, andreporter gene systems. The second part includes methods for studying components of the other major families of signal transduction: adenylyl cylase and cAMP, phosphorylated inositol lipids, phosphinositide 3-kinases, phosphlipase D and phosphatidylcholine, sphingosine kinase, and inositol1,4,5-triphosphate. Also included are chapters on baculoviral expression systems and the quantitative assay of mitogen activated protein kinases in intact cells and tissues. As with the previous edition Signal Transduction 2e covers a wide range of techniques and will be useful to both experiencedresearchers and newcomers.

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From the Publisher

Since the publication of the first edition of Signal Transduction: A Practical Approach in 1992 there has been a great deal of new information about the processes of signal transduction and consequently many new methods have been developed. This new edition has therefore been updated andextended to include the major new methods now ava...

Graeme Milligan is at University of Glasgow.
Format:PaperbackPublished:August 1, 1999Publisher:Oxford University PressLanguage:English

The following ISBNs are associated with this title:

ISBN - 10:0199637202

ISBN - 13:9780199637201

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Table of Contents

1. Direct assessment of conformational changes in G protein-coupled receptors, using fluorescene spectroscopy.1.1. Introduction1.2. Expression and purification of the v2 adrenergic receptor1.3. Fluorescence labeling and spectroscopic analysis of the v2 adrenergic receptor1.4. Site-selective fluorescent labelling of the v2 adrenergic receptor1.5. Concluding remarks2. Probing the structure of receptor-binding sites by the substituted-cysteine accessibility method2.1. Introduction2.2. Applications of the substituted-cysteine accessibility method (SCAM)2.3. Critical parameters2.4. Chemistry of SCAM2.5. Interpretation of results3. Post-translational acylation of signal transducing polypeptides: palmitoylation of G protein-coupled receptors3.1. Introduction3.2. Detection of receptor palmitoylation3.3. Kinetics of palmitoylation3.4. Assessing the palmitoylation state of the receptor at the cell surface3.5. In vitro palmitoylation of synthetic peptides4. G protein-coupled receptor phosphorylation and desensitization4.1. Introduction4.2. Techniques in the investigation of GPCR phosphorylation4.3. Identification of desensitization of phospholiphase C-coupled receptors5. G proteins and their identification5.1. Introduction5.2. Production of crude plasma membrane fractions for analysis of G proteins5.3. Gel electrophoresis of G proteins5.4. Immunological methods5.5. Quantification of G protein a subunits5.6. Functional aspects of G protein signalling6. Construction and analysis of receptor-G protein fusion proteins6.1. Introduction6.2. Methods to study G protein function6.3. Receptor-G protein fusions6.4. Expression of receptor-G protein fusions in cultured cells6.5. Assays used for functional characterization of receptor-G protein fusions6.6. Receptor-G protein fusions as research tools6.7. Summary and future perspectives7. Application of the baculoviral expression system to signal transduction7.1. Introduction7.3. The baculoviral life cycle, and constructing recombinant7.4. Recombinant protein expression and purification8. Reporter gene systems for the study of G protein-coupled receptor signal transduction in mammalian cells8.1. Introduction8.2. What is a reporter gene?8.3. Reported gene systems for the study of GPCR signal transduciton8.4. Factors influencing the design of a mammalian cell reporter-gene assay8.5. Preparation of cells for reporter-gene assays8.6. Reporter enzyme assays8.7. Reporter protein assays8.8. Summary and future perspectives9. Adenylyl cyclases and cyclic AMP9.1. Introduction9.2. Mammalian expression of recombinant Acs9.3. Measurement of intracellular cAMP level9.4. CAMP-responsive bioluminescence assays using firefly luciferase9.5 Miscellaneous systems for monitoring AC activity.

Editorial Reviews

" ... a useful addition to the bookshelf of most researchers interested in signal transduction ... it would also be benificial as a reference for other species of biologists." Cell Biology International