Using genetically engineered kinases to screen for novel protein kinase substrates: phosphorylation of kinase-associated substrates

Eblen, Scott T. ; Vinay Kumar, N. ; Weber, Michael J. (2007) Using genetically engineered kinases to screen for novel protein kinase substrates: phosphorylation of kinase-associated substrates Cold Spring Harbor Protocols, 2007 (8). No pp. given. ISSN 1940-3402

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Official URL: http://cshprotocols.cshlp.org/content/2007/4/pdb.p...

Related URL: http://dx.doi.org/10.1101/pdb.prot4638

Abstract

This protocol describes a method for detection of direct substrates of a protein kinase in cell lysates or fractions. The approach involves identification of kinase-associated substrates by immunoprecipitating a tagged form of the mutant kinase from transfected COS-1 cells and performing a kinase reaction by the addition of [γ-32P]ATP analog. This technique has been used for the phosphorylation of Extracellular signal-Regulated Kinase 2 (ERK2) substrates; however, the methodology can be applied to other protein kinases as well.

Item Type:Article
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ID Code:113511
Deposited On:25 May 2018 07:22
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