Protein Kinase Protocols

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. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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.