Hitting the target: emerging technologies in the search for kinase substrates
- PMID: 12475999
- DOI: 10.1126/stke.2002.162.pe49
Hitting the target: emerging technologies in the search for kinase substrates
Abstract
Through phosphorylation, protein kinases can alter the activity, localization, protein association, and stability of their targets. Despite the importance to our understanding of all aspects of cell biology, progress toward identifying bona fide substrates of specific protein kinases has been slow. Traditionally used techniques to identify true kinase substrates, such as genetics, yeast two-hybrid screens, and biochemical purification, are often laborious and unreliable. However, several new approaches have recently been developed and used successfully to identify genuine in vivo substrates of certain protein kinases. These methods include screening for phosphorylation of proteins from phage expression libraries, peptide library screens to determine optimal motifs favored by specific kinases, the use of phospho-motif antibodies, and an approach that uses structurally altered kinases and allele-specific adenosine triphosphate analogs and kinase inhibitors. We describe these approaches and discuss their utility and inherent caveats.
Similar articles
-
A microPLC-based approach for determining kinase-substrate specificity.Assay Drug Dev Technol. 2007 Aug;5(4):559-66. doi: 10.1089/adt.2007.072. Assay Drug Dev Technol. 2007. PMID: 17767424
-
Substrate screening of protein kinases: detection methods and combinatorial peptide libraries.Biopolymers. 2010;94(6):753-62. doi: 10.1002/bip.21506. Biopolymers. 2010. PMID: 20564046 Review.
-
PhosphoPOINT: a comprehensive human kinase interactome and phospho-protein database.Bioinformatics. 2008 Aug 15;24(16):i14-20. doi: 10.1093/bioinformatics/btn297. Bioinformatics. 2008. PMID: 18689816
-
Isolation of intrinsically active mutants of MAP kinases via genetic screens in yeast.Methods. 2006 Nov;40(3):255-61. doi: 10.1016/j.ymeth.2006.06.014. Methods. 2006. PMID: 16938468
-
Identification of protein kinase substrates by proteomic approaches.Expert Rev Proteomics. 2008 Jun;5(3):497-505. doi: 10.1586/14789450.5.3.497. Expert Rev Proteomics. 2008. PMID: 18532915 Review.
Cited by
-
Oncogenic Receptor Tyrosine Kinases Directly Phosphorylate Focal Adhesion Kinase (FAK) as a Resistance Mechanism to FAK-Kinase Inhibitors.Mol Cancer Ther. 2016 Dec;15(12):3028-3039. doi: 10.1158/1535-7163.MCT-16-0366. Epub 2016 Sep 16. Mol Cancer Ther. 2016. PMID: 27638858 Free PMC article.
-
Current technologies to identify protein kinase substrates in high throughput.Front Biol (Beijing). 2013 Apr 1;8(2):216-227. doi: 10.1007/s11515-013-1257-z. Front Biol (Beijing). 2013. PMID: 25110472 Free PMC article.
-
PPSP: prediction of PK-specific phosphorylation site with Bayesian decision theory.BMC Bioinformatics. 2006 Mar 20;7:163. doi: 10.1186/1471-2105-7-163. BMC Bioinformatics. 2006. PMID: 16549034 Free PMC article.
-
Protein kinases on carbon metabolism: potential targets for alternative chemotherapies against toxoplasmosis.Front Cell Infect Microbiol. 2023 May 23;13:1175409. doi: 10.3389/fcimb.2023.1175409. eCollection 2023. Front Cell Infect Microbiol. 2023. PMID: 37287468 Free PMC article. Review.
-
Sensitive kinase assay linked with phosphoproteomics for identifying direct kinase substrates.Proc Natl Acad Sci U S A. 2012 Apr 10;109(15):5615-20. doi: 10.1073/pnas.1119418109. Epub 2012 Mar 26. Proc Natl Acad Sci U S A. 2012. PMID: 22451900 Free PMC article.
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Other Literature Sources
Molecular Biology Databases