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Protein Kinase C Protocols

  • Book
  • © 2003

Overview

Editors:
  1. Alexandra C. Newton

    1. Department of Pharmacology, University of California at San Diego, La Jolla

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

Part of the book series: Methods in Molecular Biology (MIMB, volume 233)

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Table of contents (42 protocols)

  1. Front Matter

    Pages i-xvi
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  2. Introduction

    1. Front Matter

      Pages 1-1
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    2. The Ins and Outs of Protein Kinase C

      • Alexandra C. Newton
      Pages 3-7

    3. Early Studies of Protein Kinase C

      • Yasutomi Nishizuka, Ushio Kikkawa
      Pages 9-18

  3. Expression and Purification

    1. Front Matter

      Pages 19-19
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    2. Expression and Purification of Protein Kinase C from Insect Cells

      • Hideyuki Mukai, Yoshitaka Ono
      Pages 21-34

    3. Expression and Purification of Protein Kinase Cδ from Bacteria

      • Michael Gschwendt, Luise Stempka
      Pages 35-42

  4. Measuring the Activity of Protein Kinase C

    1. Front Matter

      Pages 43-43
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    2. Complexities in Protein Kinase C Activity Assays

      • Julianne J. Sando
      Pages 45-61

    3. Enzyme Assays for Protein Kinase C Activity

      • Julianne J. Sando, Jacquelyn K. Beals
      Pages 63-76

    4. Subcellular Translocation of Protein Kinase C

      • Julianne J. Sando, Jacquelyn K. Beals, Isa M. Hussaini
      Pages 77-85

  5. Measuring the Interaction of Protein Kinase C with Membranes

    1. Front Matter

      Pages 87-87
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    2. Measuring the Interaction of Protein Kinase C with Membranes

      • Alexandra C. Newton
      Pages 89-92

    3. Fluorescence Imaging of Protein Kinase C Translocation in Living Cells

      • Naoaki Saito
      Pages 93-103

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

      • Jennifer Giorgione, Alexandra C. Newton
      Pages 105-113

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

      • Eric A. Nalefsk, Alexandra C. Newton
      Pages 115-128

    6. [3H]Phorbol 12,13-Dibutyrate Binding Assay for Protein Kinase C and Related Proteins

      • Nancy E. Lewin, Peter M. Blumberg
      Pages 129-156

  6. Measuring the Phosphorylation of Protein Kinase C

    1. Front Matter

      Pages 157-157
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    2. Protein Kinase C Phosphorylation

      • Peter J. Parker
      Pages 159-162

    3. Pulse-Chase Analysis of Protein Kinase C

      • Mikiko Takahashi, Yoshitaka Ono
      Pages 163-170
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About this book

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).

Reviews

"...a comprehensive collection of established and emerging techniques used for the study of protein kinase C, and is quite simply an essential guide for any lab interested in cell signallin...an impressive collection of expert contributors...this is an excellent lab manual which provides very detailed and easy to follow methods and instructions that would be useful for any lab that is interested in any form of cell signalling..." - Proteomics

"...a valuable book for everybody interested in PKC, and it is a must for everybody doing research with it." - ChemBioChem

Editors and Affiliations

  • Department of Pharmacology, University of California at San Diego, La Jolla

    Alexandra C. Newton

Bibliographic Information

  • Book Title: Protein Kinase C Protocols

  • Editors: Alexandra C. Newton

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1385/1592593976

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Humana Press 2003

  • Hardcover ISBN: 978-1-58829-068-7Published: 22 May 2003

  • Softcover ISBN: 978-1-61737-335-0Published: 10 November 2010

  • eBook ISBN: 978-1-59259-397-2Published: 03 February 2008

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XVI, 584

  • Topics: Biochemistry, general

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