Membrane Proteomics: Methods and Protocols / Edition 1

by Matthew J. Peirce, Robin Wait

Overview

The membranes surrounding cells and organelles constitute their interface with the local environment. The functions of membrane proteins include cell/cell and cell/extracellular matrix recognition, the reception and transduction of extracellular signals, and the tra- port of proteins, solutes and water molecules. Abnormal membrane protein expression has profound biological effects and may, for example, underlie phenotypic and functional differences between normal and tumour cells. Moreover the accessibility, particularly of plasma proteins traversing the plasma membrane of cells, makes them of particular ut- ity to the therapeutic intervention in disease. Indeed, it is estimated that of all currently licensed pharmaceuticals, approximately 70% target proteins resident in the plasma m- brane. In theory, unbiased technologies such as proteomics have the power to define patterns of membrane protein expression characteristic of distinct states of cellular development, differentiation or disease, and thereby identify novel markers of, or targets for intervention in, disease. However, although about 25% of open reading frames in fully sequenced genomes are estimated to encode integral membrane proteins, global analysis of membrane protein expression has proved problematic. Membrane protein analysis poses unique challenges at the level of extraction, solubilization, and separation in particular, and to a lesser extent of identification and quantitation. These challenges have, however, fostered creativity, in- vation, and technical advances, many of which are brought together in Membrane P- teomics.

Product Details

ISBN-13:	9781617378898
Publisher:	Springer-Verlag New York, LLC
Publication date:	11/19/2010
Series:	Methods in Molecular Biology , #528
Edition description:	Softcover reprint of hardcover 1st ed. 2009
Pages:	342
Product dimensions:	7.60(w) x 10.24(h) x 0.03(d)

Table of Contents

In Silico Methods for Prediction of Membrane Protein Hydrophobicity and Topology.- Online Tools for Predicting Integral Membrane Proteins.- In Silico Identification of Novel G Protein Coupled Receptors.- Transcriptome-Based Identification of Candidate Membrane Proteins.- Extraction and Purification of Membrane Proteins.- Separation of Thylakoid Membrane Proteins by Sucrose Gradient Ultracentrifugation or Blue Native-SDS-PAGE Two-Dimensional Electrophoresis.- Extraction of Yeast Mihondrial Membrane Proteins by Solubilization and Detergent/Polymer Aqueous Two-Phase Partitioning.- 16-BAC/SDS-PAGE Analysis of Membrane Proteins of Yeast Mihondria Purified by Free Flow Electrophoresis.- Sequential Detergent Extraction Prior to Mass Spectrometry Analysis.- Enrichment of Brain Plasma Membranes by Affinity Two-Phase Partitioning.- Prool to Enrich and Analyze Plasma Membrane Proteins.- Proteomic Analysis of the Lymphocyte Plasma Membrane Using Cell Surface Biotinylation and Solution-Phase Isoelectric Focusing.- Identification of Target Membrane Proteins as Detected by Phage Antibodies.- Membrane Protease Degradomics: Proteomic Identification and Quantification of Cell Surface Protease Substrates.- Purification of Basolateral Integral Membrane Proteins by Cationic Colloidal Silica-Based Apical Membrane Subtraction.- Moving Closer to the Lipid Raft Proteome Using Quantitative Proteomics.- Use of Sequential Chemical Extractions to Purify Nuclear Membrane Proteins for Proteomics Identification.- Isolation of Extracellular Membranous Vesicles for Proteomic Analysis.- Separation of Membrane Proteins.- Enrichment of Human Platelet Membranes for Proteomic Analysis.- Detergents and Chaotropes for Protein Solubilization Before Two-Dimensional Electrophoresis.- Two-Dimensional Separation of Membrane Proteins by 16-BAC-SDS-PAGE.- Identification and Quantification of Membrane Proteins.- MudPIT Analysis: Application to Human Heart Tissue.- Liquid Chromatography MALDI MS/MS for Membrane Proteome Analysis.- Cysteinyl-Tagging of Integral Membrane Proteins for Proteomic Analysis Using Liquid Chromatography-Tandem Mass Spectrometry.- Quantitative Proteomics Analysis of Pancreatic Zymogen Granule Membrane Proteins.

Editorial Reviews

From the reviews:

"This is an important resource for investigators interested in understanding patterns of membrane protein expression … which are helpful in the identification of novel markers of, or targets for intervention in, disease. … It will be quite useful for research investigators and students interested in this field. … This book will be a valuable resource for investigators and students interested in learning about membrane proteomic methodologies. … High quality illustrations greatly help the understanding of the material." (Omer Iqbal, Doody’s Review Service, June, 2009)

From the Publisher

Reviewer: Omer Iqbal, MD, FACC, FESC (Loyola University Medical Center)
Description: This is an important resource for investigators interested in understanding patterns of membrane protein expression, characteristic of distinct states of cellular development, and differentiation or disease, which are helpful in the identification of novel markers of, or targets for intervention in, disease. Lucid illustrations are a unique feature of this book.
Purpose: According to the authors, this laboratory bench resource provides a comprehensive toolkit of proven methods for investigators interested in membrane proteomic methodologies. The authors are successful in achieving these worthy objectives.
Audience: The book is intended for investigators wishing to apply state-of-the-art-membrane proteomic methodologies in their research programs. It will be quite useful for research investigators and students interested in this field.
Features: Part I of the book's four parts deals with in silico methods for prediction of membrane protein hydrophobicity and topology. Part II covers extraction and purification of membrane proteins and is further divided into parts A, B and C which are subdivided into chapters. Part A explores plant membrane proteins, part B, prokaryotic membrane proteins, and part C, mammalian membrane proteins. Part III describes separation of membrane proteins and part IV covers identification and quantification of membrane proteins. The book is well illustrated with high quality, black-and-white illustrations, although color illustrations may have added further clarity.
Assessment: This book will be a valuable resource for investigators and students interested in learning about membrane proteomic methodologies. Each chapter uses the same format of abstract, introduction, materials, methods and notes followed by a list of references. High quality illustrations greatly help the understanding of the material.

Doody's Review Service