Protein Kinase Protocols

ISBN-13: 9780896037007 / Angielski / Twarda / 2000 / 365 str.

Alastair D. Reith

Opis

The wealth of primary information provided by genome sequencing projects in various species is of enormous potential value in our efforts to understand biological funtions and molecular interactions not only in normal development and cellular physiology, but also in diseases. However, utili- tion of these resources can come only from the development and application of a fully integrated set of molecular, biochemical, biophysical, and genetic skill bases. As key components of many cell signalling pathways, protein kinases are implicated in a broad variety of diseases, including cancers and neurodegenerative conditions, and offer considerable potential as tractable targets for therapeutic intervention. With these issues in mind, the present vlume has been compiled to provide examples of core skills required for ana- sis of knase-mediated signalling cascades, with particular emphasis on iden- fication of proteins according to interactive relationships and analysis of functional properties of signalling proteins. Compilation of Protein Kinase Protocols has been possible only as a - sult of the effort of all the contributors, and I am grateful to them for taking the time and having the patience to desseminate the detailied information - quired in order that others can succeed in the application of these techniques. Most importantly, I extend my deepest gratitude to Chris, Emma, and Helen for making it all worthwhile Alastair D. Reith v Contents Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . iv ix Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 Protein Kinase-Mediated Signaling Networks: Regulation and Functional Characterization Alastair D. Reith. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Recenzje

"By engaging high-ranking experts as authors of the individual chapters the editor has succeeded in compiling an updated an highly useful compendium of today's technology in protein kinase research. besides its topicality, this book has the particular advantage that the chapters are arranged according to a constantly recurring scheme, that is, beginning with a comprehensive introduction, which provides the appropriate key references, and then continuing with very detailed experimental instructions, which are given clearly enough to enable even a less experienced experimentator to follow. The "Notes" at the end of each chapter are extremely helpful, in which the experimental steps are critically examined and explained in detail....for students and newcomers in the field it provides an excellent introduction to the methodology of protein kinase research, and for the experienced scientist it will be a highly valuable work of reference and an indispensable companion in daily laboratory work." - Chembiochem

"The authors are all experienced researchers with some eminent names among them. Each of the chapters has been contributed by scientists with considerable expertise in the areas included, leaving the reader with knowledge that the protocols are thoroughly tried and tested. All in all, the book presents a wide range of specialized techniques which those working on (or intending to work on) kinases would find a very useful addition to their laboratories." - Cell Biology International

Spis treści

Protein Kinase-Mediated Signaling Networks: Regulation and Functional Characterization, Alastair D. Reith. Cloning Protein Tyrosine Kinases by Screening cDNA Libraries with Antiphosphotyrosine Antibodies, Lisa D. Chong and Ira O. Daar. Immunoprecipitation and Western Blotting of Phosphotyrosine-Containing Proteins, Kathleen M. Woods-Ignatoski. Two-Dimensional Phosphoamino Acid Analysis, Peter Blume-Jensen and Tony Hunter. Two-Dimensional Phosphopeptide Mapping of Receptor Tyrosine Kinases, Jeremy M. Tavaré and Tarik Issad. Identification of the Sites of Phosphorylation in Proteins Using High Performance Liquid Chromatography and Mass Spectrometry, A. Grey Craig. Phosphorylation of Smad Signaling Proteins by Receptor Serine/ Threonine Kinases, Serhiy Souchelnytskyi, Lars Rönnstrand, Carl-Henrik Heldin, and Peter ten Dijke. Assays for Mitogen-Activated Protein Kinase (MAPK) Subtypes and MAPK Activating Protein Kinase-2 (MAPKAP K-2) Using a Common Cell Lysate, Rozen Le Panse, Navita Rampersaud, and Louis C. Mahadevan. JAK-Mediated Phosphorylation and Activation of STAT Signaling Proteins: Analysis by Phosphotyrosine Blotting and EMSA, Nicola Broughton and Mark S. Burfoot. Assays for Glycogen Synthase Kinase-3 (GSK-3), Darren Cross. Cyclin-Dependent Kinases and Cyclin-Dependent Kinase Inhibitors: Detection Methods and Activity Measurements, Gavin Brooks. Protein Histidine Kinase, Harry R. Matthews and Karina Chan. Cloning and Expression of Recombinant HEXA-HIS Tagged ZAP-70 Using the Baculovirus System, Tracey Purton, Sandra Wilkinson, and Edward J. Murray. Analysis of Protein Kinase Subcellular Localization by Visualization of GFP Fusion Proteins, Ralph A. Zirngibl and Peter Greer. Detection of Phosphorylation-Dependent Interactions by Far-Western Gel Overlay, Mary Rose Burnham, Regina DeBerry, and Amy H. Bouton. Purification of Tyrosine-Phosphorylated Proteins by Immunoaffinity Chromatography and Direct Cloning of Their cDNAs from Bacterial Expression Libraries,Pier Paolo Di Fiore and Francesca Fazioli. Identification of Receptor Tyrosine Kinase (RTK) Substrates by the Cloning of Receptor Targets (CORT) Strategy, Roger J. Daly. Identification of Receptor Tyrosine Kinase Associating Proteins Using the Yeast Two-Hybrid System, Sally A. Prigent. cDNA Expression Cloning and Charaterization of Phosphorylation Dependent Protein Interactors Using the Yeast Tribrid System, Christoph Volpers, Manuel Lubinus, Mark A. Osborne, and Jarema P. Kochan. Analysis of SH2 Domain-Phosphopeptide Interactions by Isothermal Titration Calorimetry and Surface Plasmon Resonance, George Panayotou and John Ladbury. Cloning and Characterization of RTK Ligands Using Receptor-Alkaline Phosphatase Fusion Proteins, Hwai-Jong Cheng and John G. Flanagan. Isolation and Characterization of 'Orphan-RTK' Ligands Using an Integrated Biosensor Approach, Martin Lackmann. Index.

Blurb

As key components of many cell signaling pathways, protein kinases are implicated in a broad variety of diseases, including cancers and neurodegenerative conditions, and offer considerable potential as tractable targets for therapeutic intervention. In Protein Kinase Protocols, a panel of highly skilled laboratory investigators describe both basic and more sophisticated methods for the analysis of kinase-mediated signaling cascades, with emphasis on the identification of proteins according to their interactive relationships and the analysis of their functional properties. Described in step-by-step detail, these readily reproducible techniques offer novices quick access to a complicated field, and provide more experienced investigators many novel time-saving ploys. Emphasis is given to the critical technical steps that are often omitted from methods published in the primary literature. There are also tips on potential pitfalls and copious notes on how to adjust the protocols to work in related systems.
Broad in its range of techniques and thoroughly detailed to help ensure experimental success, Protein Kinase Protocols offers both novice and experienced investigators powerful tools for understanding the functional roles of specific protein kinases within signaling cascades and for identification and evaluation of novel therapeutic targets.