Journal article
Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach
Journal of visualized experiments, (144)
21 Feb 2019
PMID: 30855574
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
The study of kinase-substrate relationships is essential to gain a complete understanding of the functions of these enzymes and their downstream targets in both physiological and pathological states. CK2 is an evolutionarily conserved serine/threonine kinase with a growing list of hundreds of substrates involved in multiple cellular processes. Due to its pleiotropic properties, identifying and characterizing a comprehensive set of CK2 substrates has been particularly challenging and remains a hurdle in the study of this important enzyme. To address this challenge, we have devised a versatile experimental strategy that enables the targeted enrichment and identification of putative CK2 substrates. This protocol takes advantage of the unique dual co-substrate specificity of CK2 allowing for specific thiophosphorylation of its substrates in a cell or tissue lysate. These substrate proteins are subsequently alkylated, immunoprecipitated, and identified by liquid chromatography/tandem mass spectrometry (LC-MS/MS). We have previously used this approach to successfully identify CK2 substrates from Drosophila ovaries and here we extend the application of this protocol to human glioblastoma cells, illustrating the adaptability of this method to investigate the biological roles of this kinase in various model organisms and experimental systems.
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Details
- Title
- Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach
- Creators
- John E. Chojnowski - Drexel UniversityEmily A. McMillan - Drexel UniversityTodd Strochlic - Drexel University
- Publication Details
- Journal of visualized experiments, (144)
- Publisher
- Journal Of Visualized Experiments
- Number of pages
- 8
- Grant note
- Commonwealth Universal Research Enhancement grant from the Pennsylvania Department of Health
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology
- Web of Science ID
- WOS:000462905000049
- Scopus ID
- 2-s2.0-85062718436
- Other Identifier
- 991020837830304721
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InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Biochemistry & Molecular Biology