Cover image for Membrane proteomics : methods and protocols
Title:
Membrane proteomics : methods and protocols
Series:
Methods in molecular biology, 528
Publication Information:
New York; Humana Press, 2009
Physical Description:
xiii, 342 p. : ill. ; 27 cm.
ISBN:
9781603273091

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30000010194927 QP552.M44 M454 2009 Open Access Book Book
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Summary

Summary

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 de?ne 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 identi?cation and quantitation. These challenges have, however, fostered creativity, in- vation, and technical advances, many of which are brought together in Membrane P- teomics.


Table of Contents

Henry Bigelow and Burkhard RostMatthew N. Davies and Darren R. FlowerEdward J. Evans and Lawrence Hene and Mai Vuong and S. Hussain I. Abidi and Simon J. DavisGian Maria D'Amici and Christian G. Huber and Lello ZollaHenrik Everberg and Niklas Gustavsson and Folke TjerneldRalf J. Braun and Norbert Kinkl and Hans Zischka and Marius UeffingFiona M. McCarthy and Amanda M. Cooksey and Shane C. BurgessJens Schindler and Hans Gerd NothwangJacek R. WiśniewskiMatthew J. Peirce and Andrew P. Cope and Robin WaitCecile A.W. Geuijen and Arjen Q. Bakker and John de KruifGeorgina S. Butler and Richard A. Dean and Derek Smith and Christopher M. OverallRobert J.A. Goode and Richard J. SimpsonLeonard J. FosterNadia Korfali and Elizabeth A. L. Fairley and Selene K. Swanson and Laurence Florens and Eric C. SchirmerRommel A. Mathias and Justin W. &Lbar;im and Hong Ji and Richard J. SimpsonDavid W. Greening and Kristen M. Glenister and Rosemary L. Sparrow and Richard J. SimpsonThierry RabilloudHans Gerd Nothwang and Jens SchindlerKelli G. Kline and Christine C. WuNan Wang and J. Bryce Young and Liang LiSrijeet K. Mitra and Michael B. GosheXuequn Chen and Philip C. Andrews
Prefacep. v
Contributorsp. xi
Part I In Silico Methods for Prediction of Membrane Protein Hydrophobicity and Topology
1 Online Tools for Predicting Integral Membrane Proteinsp. 3
2 In Silico Identification of Novel G Protein Coupled Receptorsp. 25
3 Transcriptome-Based Identification of Candidate Membrane Proteinsp. 37
Part II Extraction and Purification of Membrane Proteins
Part A Plant Membrane Proteins
4 Separation of Thylakoid Membrane Proteins by Sucrose Gradient Ultracentrifugation or Blue Native-SDS-Page Two-Dimensional Electrophoresisp. 61
Part B Prokaryotic Membrane Proteins
5 Extraction of Yeast Mitochondrial Membrane Proteins by Solubilization and Detergent/Polymer Aqueous Two-Phase Partitioningp. 73
6 16-BAC/SDS-Page Analysis of Membrane Proteins of Yeast Mitochondria Purified by Free Flow Electrophoresisp. 83
Part C Mammalian Membrane Proteins
7 Sequential Detergent Extraction Prior to Mass Spectrometry Analysisp. 111
8 Enrichment of Brain Plasma Membranes by Affinity Two-Phase Partitioningp. 119
9 Protocol to Enrich and Analyze Plasma Membrane Proteinsp. 127
10 Proteomic Analysis of the Lymphocyte Plasma Membrane Using Cell Surface Biotinylation and Solution-Phase Isoelectric Focusingp. 135
11 Identification of Target Membrane Proteins as Detected by Phage Antibodiesp. 141
12 Membrane Protease Degradomics: Proteomic Identification and Quantification of Cell Surface Protease Substratesp. 159
13 Purification of Basolateral Integral Membrane Proteins by Cationic Colloidal Silica-Based Apical Membrane Subtractionp. 177
14 Moving Closer to the Lipid Raft Proteome Using Quantitative Proteomicsp. 189
15 Use of Sequential Chemical Extractions to Purify Nuclear Membrane Proteins for Proteomics Identificationp. 201
16 Isolation of Extracellular Membranous Vesicles for Proteomic Analysisp. 227
Part III Separation of Membrane Proteins
17 Enrichment of Human Platelet Membranes for Proteomic Analysisp. 245
18 Detergents and Chaotropes for Protein Solubilization Before Two-Dimensional Electrophoresisp. 259
19 Two-Dimensional Separation of Membrane Proteins by 16-BAC-SDS-Pagep. 269
Part IV Identification and Quantification of Membrane Proteins
20 MudPIT Analysis: Application to Human Heart Tissuep. 281
21 Liquid Chromatography Maldi MS/MS for Membrane Proteome Analysisp. 295
22 Cysteinyl-Tagging of Integral Membrane Proteins for Proteomic Analysis Using Liquid Chromatography-Tandem Mass Spectrometryp. 311
23 Quantitative Proteomics Analysis of Pancreatic Zymogen Granule Membrane Proteinsp. 327
Indexp. 339