Protein kinase C protocols
Author(s)
Bibliographic Information
Protein kinase C protocols
(Methods in molecular biology / John M. Walker, series editor, v. 233)
Humana Press, c2003
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Note
Includes bibliographical references and index
Description and Table of Contents
Description
Since the discovery that protein kinase C (PKC) transduces the ab- dance of signals that result in phospholipid hydrolysis, this enzyme has been at the forefront of research in signal transduction. Protein Kinase C Protocols covers fundamental methods for studying the structure, function, regulation, subcellular localization, and macromolecular interactions of PKC. Protein Kinase C Protocols is divided into 11 sections representing the major aspects of PKC regulation and function. Part I contains an introduction and a historical perspective on the discovery of PKC by Drs. Yasutomi Nishizuka and Ushio Kikkawa. Part II describes methods to purify PKC. Part III describes the standard methods for measuring PKC activity: its enzymatic activity and its stimulus-dependent translocation from the cytosol to the membrane. Part IV describes methods for measuring the membrane interaction of PKC in vivo and in vitro. Part V provides methodologies and techniques for measuring the ph- phorylation state of PKC, including a protocol for measuring the activity of PKC's upstream kinase, PDK-1. Novel methods for identifying substrates are described in Part VI. Part VII presents protocols for expressing and analyzing the membrane targeting domains of PKC. Part VIII provides a comprehensive c- pilation of methods used to identify binding partners for PKC. Part IX describes pharmacological probes used to study PKC. The book ends with a presentation of genetic approaches to study PKC (Part X) and a discussion of approaches used to study PKC in disease (Part XI).
Table of Contents
Part I. Introduction
The Ins and Outs of Protein Kinase C
Alexandra C. Newton
Early Studies of Protein Kinase C: A Historical Perspective
Yasutomi Nishizuka and Ushio Kikkawa
Part II. Expression and Purification
Expression and Purification of Protein Kinase C from Insect Cells
Hideyuki Mukai and Yoshitaka Ono
Expression and Purification of Protein Kinase Cd from Bacteria
Michael Gschwendt and Luise Stempka
Part III. Measuring the Activity of Protein Kinase C
Complexities in Protein Kinase C Activity Assays: An Introduction
Julianne J. Sando
Enzyme Assays for Protein Kinase C Activity
Julianne J. Sando and Jacquelyn K. Beals
Subcellular Translocation of Protein Kinase C
Julianne J. Sando, Jacquelyn K. Beals, and Isa M. Hussaini
Part IV. Measuring the Interaction of Protein Kinase C with Membranes
Measuring the Interaction of Protein Kinase C with Membranes: An Introduction
Alexandra C. Newton
Fluorescence Imaging of Protein Kinase C Translocation in Living Cells
Naoaki Saito
Measuring the Binding of Protein Kinase C to Sucrose-Loaded Vesicles
Jennifer Giorgione and Alexandra C. Newton
Use of Stopped-Flow Fluorescence Spectroscopy to Measure Rapid Membrane Binding by Protein Kinase C
Eric A. Nalefski and Alexandra C. Newton
[3H]Phorbol 12,13-Dibutyrate Binding Assay for Protein Kinase C and Related Proteins
Nancy E. Lewin and Peter M. Blumberg
Part V. Measuring the Phosphorylation of Protein Kinase C
Protein Kinase C Phosphorylation: An Introduction
Peter J. Parker
Pulse-Chase Analysis of Protein Kinase C
Mikiko Takahashi and Yoshitaka Ono
PDK-1 and Protein Kinase C Phosphorylation
Alex Toker
In Vitro Autophosphorylation of Protein Kinase C Isozymes
Antonio M. Pepio and Wayne S. Sossin
Tyrosine Phosphorylation of Protein Kinase C
Toshiyoshi Yamamoto, Emiko Yamauchi, Hisaaki Taniguchi, Hidenori Matsuzaki, and Ushio Kikkawa
Methods to Study Dephosphorylation of Protein Kinase C In Vivo
Karen England and Martin Rumsby
Phosphopeptide-Specific Antibodies to Protein Kinase C
Wayne S. Sossin
Part VI. Identifying Protein Kinase C Substrates
Identifying Protein Kinase C Substrates: An Introduction
Alex Toker
A Chemical Genetic Approach for the Identification of Direct Substrates of Protein Kinases
Kavita Shah and Kevan M. Shokat
Studying the Optimal Peptide Substrate Motifs of Protein Kinase C Using Oriented Peptide Libraries
Michael B. Yaffe
Part VII. Structural Analysis of Protein Kinase C
Structural Analysis of Protein Kinase C: An Introduction
James H. Hurley
Bacterial Expression and Purification of C1 and C2 Domains of Protein Kinase C Isoforms
Wonhwa Cho, Michelle Digman, Bharath Ananthanarayanan, and Robert V. Stahelin
Crystallization of the Protein Kinase Cd C1B Domain
Gongyi Zhang and James H. Hurley
Part VIII. Methods for Detecting Binding Proteins
Methods for Detecting Binding Proteins: An Introduction
Gerda Endemann and Daria Mochly-Rosen
Detection of Protein Kinase-Binding Partners by the Yeast Two-Hybrid Analysis
Betty Y. Chang and Christine A. Cartwright
Glutathione S-Transferase Pull-Down Assay
Deborah Schechtman, Daria Mochly-Rosen, and Dorit Ron
Overlay Method for Detecting Protein-Protein Interactions
Deborah Schechtman, Christopher Murriel, Rachel Bright, and Daria Mochly-Rosen
An Overlay Assay for Detecting Protein Kinase C-Binding Proteins and Substrates
Susan Jaken
Functional Proteomic Analysis of the Protein Kinase C Signaling System
Jason M. Pass, Jun
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