Protein Kinase C Protocols / edited by Alexandra C. Newton.

Format:

Book

Contributor:

Newton, Alexandra C., editor.

SpringerLink (Online service)

Series:

Methods in molecular biology 1064-3745 ; 233.

Springer Protocols (Springer-12345)

Methods in Molecular Biology™, 1064-3745 ; 233

Language:

English

Subjects (All):

Life sciences.

Biochemistry.

Life Sciences.

Biochemistry, general.

Local Subjects:

Life Sciences.

Biochemistry, general.

Physical Description:

1 online resource (XVI, 584 pages).

Contained In:

Springer eBooks

Place of Publication:

Totowa, NJ : Humana Press, 2003.

System Details:

text file PDF

Summary:

Protein kinase C impacts on nearly every aspect of signal transduction, placing it at the forefront of research in how cells transduce information. In Protein Kinase C Protocols, leading investigators from academia and industry present a comprehensive collection of classical and emerging laboratory techniques for the study of this influential protein, all optimized for successful and reproducible results. Described in step-by-step detail, these methods can be easily used to explore the structure, function, regulation, subcellular localization, and macromolecular interactions of protein kinase C. Included in this collection are easy-to-follow techniques to purify protein kinase C, to measure its activity and its membrane interaction in vivo and in vitro, and to identify substrates. There are also methodologies and techniques for measuring the phosphorylation state of protein kinase C, for expressing and analyzing the membrane-targeting domains of protein kinase C, and for detecting binding proteins. Additional chapters address pharmacological probes, genetic approaches, and protein kinase C in disease. Each protocol is introduced in the context of protein kinase C function and regulation and contains many notes on how best to deal with the problems that may occur. Comprehensive and authoritative, Protein Kinase C Protocols offers a timely compilation of biophysical, biochemical, cell biological, and molecular biological approaches that will ensure successful results in every laboratory working in this rapidly advancing field.

Contents:

The Ins and Outs of Protein Kinase C

Early Studies of Protein Kinase C

Expression and Purification

Expression and Purification of Protein Kinase C from Insect Cells

Expression and Purification of Protein Kinase C? from Bacteria

Measuring the Activity of Protein Kinase C

Complexities in Protein Kinase C Activity Assays

Enzyme Assays for Protein Kinase C Activity

Subcellular Translocation of Protein Kinase C

Measuring the Interaction of Protein Kinase C with Membranes

Fluorescence Imaging of Protein Kinase C Translocation in Living Cells

Measuring the Binding of Protein Kinase C to Sucrose-Loaded Vesicles

Use of Stopped-Flow Fluorescence Spectroscopy to Measure Rapid Membrane Binding by Protein Kinase C

[3H]Phorbol 12,13-Dibutyrate Binding Assay for Protein Kinase C and Related Proteins

Measuring the Phosphorylation of Protein Kinase C

Protein Kinase C Phosphorylation

Pulse-Chase Analysis of Protein Kinase C

PDK-1 and Protein Kinase C Phosphorylation

In Vitro Autophosphorylation of Protein Kinase C Isozymes

Tyrosine Phosphorylation of Protein Kinase C

Methods to Study Dephosphorylation of Protein Kinase C In Vivo

Phosphopeptide-Specific Antibodies to Protein Kinase C

Identifying Protein Kinase C Substrates

A Chemical Genetic Approach for the Identification of Direct Substrates of Protein Kinases

Studying the Optimal Peptide Substrate Motifs of Protein Kinase C Using Oriented Peptide Libraries

Structural Analysis of Protein Kinase C

Bacterial Expression and Purification of C1 and C2 Domains of Protein Kinase C Isoforms

Crystallization of the Protein Kinase C? C1B Domain

Methods for Detecting Binding Proteins

Detection of Protein Kinase-Binding Partners by the Yeast Two-Hybrid Analysis

Glutathione S-Transferase Pull-Down Assay

Overlay Method for Detecting Protein-Protein Interactions

An Overlay Assay for Detecting Protein Kinase C-Binding Proteins and Substrates

Functional Proteomic Analysis of the Protein Kinase C Signaling System

Pharmacological Probes for Protein Kinase C

Applications of Inhibitors for Protein Kinase C and Their Isoforms

Phorbol Esters as Probes for the Study of Protein Kinase C Function

Irreversible Inactivation of Protein Kinase C Isozymes by Thiol-Reactive Peptide Substrate Analogs

Genetic Approaches to Studying Protein Kinase C

Animal Models in the Study of Protein Kinase C Isozymes

Genetic Manipulation of Protein Kinase C In Vivo

Yeast as a Host to Screen for Modulators and Regulatory Regions of Mammalian Protein Kinase C Isoforms

Protein Kinase C in Disease

Characterization of the Role of Protein Kinase C Isozymes in Colon Carcinogenesis Using Transgenic Mouse Models

Alcohol Addiction.

Other Format:

Printed edition:

ISBN:

9781592593972

Access Restriction:

Restricted for use by site license.