Protein Kinase Protocols

Buch | Softcover

365 Seiten

2010 | Softcover reprint of hardcover 1st ed. 2001
Humana Press Inc. (Verlag)
978-1-61737-141-7 (ISBN)

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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...

Protein Kinase-Mediated Signaling Networks.- Cloning Protein Tyrosine Kinases by Screening cDNA Libraries with Antiphosphotyrosine Antibodies.- Immunoprecipitation and Western Blotting of Phosphotyrosine-Containing Proteins.- Two-Dimensional Phosphoamino Acid Analysis.- Two-Dimensional Phosphopeptide Mapping of Receptor Tyrosine Kinases.- Identification of the Sites of Phosphorylation in Proteins Using High Performance Liquid Chromatography and Mass Spectrometry.- Phosphorylation of Smad Signaling Proteins by Receptor Serine/Threonine Kinases.- Assays for Mitogen-Activated Protein Kinase (MARK) Subtypes and MARK Activating Protein Kinase-2 (MAPKAP K-2) Using a Common Cell Lysate.- JAK-Mediated Phosphorylation and Activation of STAT Signaling Proteins.- Assays for Glycogen Synthase Kinase-3(GSK-3).- Cyclin-Dependent Kinases and Cyclin-Dependent Kinase Inhibitors.- Protein Histidine Kinase.- Cloning and Expression of Recombinant HEXA-HIS Tagged ZAP-70 Using the Baculovirus System.- Analysis of Protein Kinase Subcellular Localization by Visualization of GFP Fusion Proteins.- Detection of Phosphorylation-Dependent Interactions by Far-Western Gel Overlay.- Purification of Tyrosine-Phosphorylated Proteins by Immunoaffinity Chromatography and Direct Cloning of Their cDNAs from Bacterial Expression Libraries.- Identification of Receptor Tyrosine Kinase (RTK) Substrates by the Cloning of ReceptorTargets (CORT) Strategy.- Identification of Receptor Tyrosine Kinase Associating Proteins Using the Yeast Two-Hybrid System.- cDNA Expression Cloning and Characterization of Phosphorylation Dependent Protein Interactors Using the Yeast Tribrid System.- Analysis of SH2 Domain - Phosphopeptide Interactions by Isothermal Titration Calorimetry and Surface Plasmon Resonance.- Cloning and Characterization of RTK Ligands Using Receptor-Alkaline Phosphatase Fusion Proteins.- Isolation and Characterization of “Orphan-RTK” Ligands Using an Integrated Biosensor Approach.

Erscheint lt. Verlag	10.11.2010
Reihe/Serie	Methods in Molecular Biology ; 124
Zusatzinfo	XIII, 365 p.
Verlagsort	Totowa, NJ
Sprache	englisch
Maße	152 x 229 mm
Themenwelt	Naturwissenschaften ► Biologie ► Biochemie
ISBN-10	1-61737-141-6 / 1617371416
ISBN-13	978-1-61737-141-7 / 9781617371417
Zustand	Neuware