Identification and Expression of G Protein-Coupled Receptors / Edition 1

by Kevin R. Lynch

Overview

The past decade has seen tremendous advances in the study of G protein-coupled receptors (GPCRs), including the molecular cloning and identification of more than 100 hundred GPCR genes. But while GPCRs serve as targets for more than 300 medicines in the modern pharmacopoeia, the shrinking pool of known ligands and the continuing discovery of orphan GPCR genes have underscored the need for new approaches to ligand identification.

Identification and Expression of G Protein-Coupled Receptors addresses this new direction in GPCR biochemistry-offering a definitive laboratory bench manual that emphasizes expression over primary cloning strategies. In a series of expert contributions by well-known researchers, this book provides detailed protocols for various expression systems-from bacteria to mammalian cells-as well as straightforward opinions on the advantages and shortcomings of each approach. Topics covered include:
* Homology screening and the polymerase chain reaction in the cloning of GPCR genes
* Cloning of GPCRs using mammalian cell expression
* GPCR informatics and the orphan problem
* The use of Xenopus laevis oocytes for the study of GPCRs
* Stable expression of GPCRs in mammalian cells
* Heterologous expression in primary cell cultures
* Expression of GPCR in Escherichia coli
* Large scale expression and purification of GPCRs in mammalian cells
* High-level expression of GPCRs in the Baculovirus/Sf9 cell expression system
* Expression of GPCRs in Drosophila Schneider 2 cells
* Methods for genetic analysis and ligand identification using heterologous GPCRs expressed in Saccharomyces cerevisiae

Supplemented with numerous photographs and illustrations, Identification and Expression of G Protein-Coupled Receptors is important reading for biochemists, pharmacologists, neuroscientists, structural biologists, and anyone involved in GPCR-based research. It delivers a wealth of useful advice, practical tips, and invaluable insight into trends at the cutting-edge of current research.

Product Details

ISBN-13:	9780471194934
Publisher:	Wiley
Publication date:	05/10/1999
Series:	Receptor Biochemistry and Methodology , #21
Pages:	222
Product dimensions:	7.48(w) x 9.92(h) x 0.47(d)

About the Author

Kevin R. Lynch is the author of Identification and Expression of G Protein-Coupled Receptors, published by Wiley.

Table of Contents

Cloning of G Protein-Coupled Receptor Genes: The Use of Homology Screening and the Polymerase Chain Reaction (A. Marchese, et al.).

Practical Approaches for Cloning G Protein-Coupled Receptors Using Mammalian Cell Expression (T. Murphy).

G Protein-Coupled Receptor Informatics and the Orphan Problem (K. Lynch).

The Use of Xenopus laevis Oocytes for the Study of G Protein-Coupled Receptors (A. Lee & M. Durieux).

Stable Expression of G Protein-Coupled Receptors in Mammalian Cells (K. Neve & R. Neve).

Heterologous Expression in Primary Cell Cultures (R. Leonard).

Expression of G Protein-Coupled Receptors in Escherichia coli (R. Grisshammer).

Large-Scale Expression/Purification of G Protein-Coupled Receptors Mammalian Cells (A. Robeva & J. Linden).

High-Level Expression of G Protein-Coupled Receptors in the Baculovirus/SF9 Cell Expression System (N. Rondeau, et al.).

Expression of G Protein-Coupled Receptors in Drosophila Schneider 2 Cells (M. Graziano, et al.).

Heterologous G Protein-Coupled Receptors Expressed in Saccharomyces cerevisiae: Methods for Genetic Analysis and Ligand Identification (M. Pausch, et al.).

Index.

Editorial Reviews

Reviewer: Eugene A Davidson, PhD (Georgetown University School of Medicine)
Description: This is a guide to the identification and expression of G protein-coupled receptors.
Purpose: This volume is intended as a "how-to" laboratory accessory for investigators wanting to express G protein-coupled receptors.
Audience: Investigators with a need to express such receptors are the intended audience, but they will have difficulty in decision making since there is a lack of critical commentary on the pros and cons of the various technologies.
Features: The coverage is reasonably complete in that the most common methods of protein expression are described — bacteria, mammalian cells, insect cells, oocytes, and yeast. Attention is also given to the growing problem of information management and there is an extensive listing of cloned receptors together with accession data.
Assessment: It is not uncommon for a given protein to be effectively expressed by one method (e.g., yeast) and fail miserably in a different genetic background. Given a new candidate, how does an investigator decide on the best strategic approach? If one is willing to try several, this volume will have value. If resources are limited so that the shotgun approach is not feasible, prudent consultation is likely to provide a better outcome. It also should be noted that several of the chapters provide descriptive material as opposed to protocols. In this regard, the utility of the book as a laboratory adjunct is limited.

Doody's Review Service

1 Star from Doody

This is a guide to the identification and expression of G protein-coupled receptors. This volume is intended as a ""how-to"" laboratory accessory for investigators wanting to express G protein-coupled receptors. Investigators with a need to express such receptors are the intended audience, but they will have difficulty in decision making since there is a lack of critical commentary on the pros and cons of the various technologies. The coverage is reasonably complete in that the most common methods of protein expression are described—bacteria, mammalian cells, insect cells, oocytes, and yeast. Attention is also given to the growing problem of information management and there is an extensive listing of cloned receptors together with accession data. It is not uncommon for a given protein to be effectively expressed by one method (e.g., yeast) and fail miserably in a different genetic background. Given a new candidate, how does an investigator decide on the best strategic approach? If one is willing to try several, this volume will have value. If resources are limited so that the shotgun approach is not feasible, prudent consultation is likely to provide a better outcome. It also should be noted that several of the chapters provide descriptive material as opposed to protocols. In this regard, the utility of the book as a laboratory adjunct is limited.

Eugene A. Davidson

A laboratory bench manual for biochemists, pharmacologists, neuro- scientists, structural biologists, and other investing the receptors. Emphasizing expression over primary cloning strategies, provides detailed protocols for various expression systems from bacteria to mammalian cells, and offers opinions on the advantages and shortcoming of each approach. Among the topics are homology screening and the polymerase chain reaction in cloning the receptor genes, heterologous expression in primary cell cultures, large-scale expression and purification in mammalian cells, Schneider 2 cells, and methods for genetic analysis and ligand identification using heterologous receptors expressed in . Annotation c. by Book News, Inc., Portland, Or.

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