Protein-Protein Interactions : Methods and Applications / edited by Haian Fu.

Format:

Book

Contributor:

Fu, Haian, editor.

SpringerLink (Online service)

Series:

Methods in molecular biology 1064-3745 ; 261.

Springer Protocols (Springer-12345)

Methods in Molecular Biology, 1064-3745 ; 261

Language:

English

Subjects (All):

Life sciences.

Biochemistry.

Life Sciences.

Biochemistry, general.

Local Subjects:

Life Sciences.

Biochemistry, general.

Physical Description:

1 online resource (XVI, 532 pages).

Contained In:

Springer eBooks

Place of Publication:

Totowa, NJ : Humana Press, 2004.

System Details:

text file PDF

Summary:

Proteins continuously interact with each other to determine cell fate. Consequently, an examination of just when such protein-protein interactions occur and how they are controlled is essential for understanding the molecular mechanism of biological processes, elucidating the molecular basis of diseases, and identifying potential targets for therapeutic interventions. In Protein-Protein Interactions: Methods and Applications, leading experts describe in detail their highly successful biochemical, biophysical, genetic, and computational techniques for studying these interactions. Their readily reproducible methods demonstrate how to identify protein interaction partners, qualitatively or quantitatively measure protein-protein interactions, monitor protein-protein interactions as they occur in living cells, and determine interaction interfaces. The techniques described utilize a variety of cutting-edge technologies, including surface plasmon resonance (SRP), fluorescence resonance energy transfer (FRET), fluorescence polarization (FP), isothermal titration calorimetry (ITC), circular dichroism (CD), protein fragment complementation assays (PCA), various two-hybrid systems, and proteomics- and bioinformatics-based approaches, such as the Scansite program for computational analysis. Each time-tested protocol includes a background introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Authoritative and highly practical, Protein-Protein Interactions: Methods and Applications offers both beginning and experienced investigators a full range of the powerful tools needed for deciphering how proteins interact to form biological networks, as well as for unraveling protein-protein interactions in disease in the search for novel therapeutic targets.

Contents:

Overview

Structural Basis of Protein-Protein Interactions

Quantitative Analysis of Protein-Protein Interactions

In Vitro Techniques

Characterization of Protein-Protein Interactions by Isothermal Titration Calorimetry

Circular Dichroism Analysis for Protein-Protein Interactions

Protein-Protein Interaction Analysis by Nuclear Magnetic Resonance Spectroscopy

Measuring Rhodopsin-G-Protein Interactions by Surface Plasmon Resonance

Using Light Scattering to Determine the Stoichiometry of Protein Complexes

Sedimentation Equilibrium Studies

Analysis of Protein-Protein Interactions by Simulation of Small-Zone Gel Filtration Chromatography

Fluorescence Gel Retardation Assay to Detect Protein-Protein Interactions

Fluorescence Polarization Assay to Quantify Protein-Protein Interactions

Studying Protein-Protein Interactions via Blot Overlay or Far Western Blot

Glutathione-S-Transferase-Fusion Based Assays for Studying Protein-Protein Interactions

Affinity Capillary Electrophoresis Analyses of Protein-Protein Interactions in Target-Directed Drug Discovery

Mapping Protein-Ligand Interactions by Hydroxyl-Radical Protein Footprinting

Use of Phage Display and Polyvalency to Design Inhibitors of Protein-Protein Interactions

Detecting Protein-Protein Interactions in Heterologous Systems

A Bacterial Two-Hybrid System Based on Transcription Activation

Using the Yeast Two-Hybrid System to Identify Interacting Proteins

Analysis of Protein-Protein Interactions Utilizing Dual Bait Yeast Two-Hybrid System

The Split-Ubiquitin Membrane-Based Yeast Two-Hybrid System

Reverse Two-Hybrid Techniques in the Yeast Saccharomyces cerevisiae

Mammalian Two-Hybrid Assay for Detecting Protein-Protein Interactions In Vivo

Co-Immunoprecipitation from Transfected Cells

Probing Protein-Protein Interactions in Living Cells

Microscopic Analysis of Fluorescence Resonance Energy Transfer (FRET)

Monitoring Molecular Interactions in Living Cells Using Flow Cytometric Analysis of Fluorescence Resonance Energy Transfer

Fluorescence Correlation Spectroscopy

Confocal Microscopy for Intracellular Co-Localization of Proteins

Mapping Biochemical Networks With Protein-Fragment Complementation Assays

In Vivo Protein Cross-Linking

Proteomics-Based Approaches

Computational Prediction of Protein-Protein Interactions

Affinity Methods for Phosphorylation-Dependent Interactions

Two-Dimensional Gel Electrophoresis for Analysis of Protein Complexes

Sample Preparation of Gel Electrophoretically Separated Protein Binding Partners for Analysis by Mass Spectrometry

Quantitative Protein Analysis by Solid Phase Isotope Tagging and Mass Spectrometry

Internet Resources for Studying Protein-Protein Interactions.

Other Format:

Printed edition:

ISBN:

9781592597628

Access Restriction:

Restricted for use by site license.