Substrate screening of protein kinases: detection methods and combinatorial peptide libraries
- PMID: 20564046
- DOI: 10.1002/bip.21506
Substrate screening of protein kinases: detection methods and combinatorial peptide libraries
Abstract
The study of protein kinases has become a matter of great importance in the development of new drugs for the treatment of diseases, including cancer and inflammation. Substrate screening is the first step in the fundamental investigation of protein kinases and the development of inhibitors for use in drug discovery. Towards this goal, various studies have been reported regarding the development of phospho-peptide detection methods and the screening of phosphorylated peptide sites by protein kinases. This review introduces the detection methods for phosphorylation events using the reagents with (γ(32)P)ATP, ligand-linked ATP, phospho-peptide-specific antibodies and metal chelating compounds. Chemical modification methods using β-elimination for the detection of phospho-Ser/Thr peptides are introduced as well. In addition, the implementations of combinatorial peptide libraries for screening peptide substrates of protein kinases are discussed. The phage display approach has been suggested as an alternative method of using synthetic peptides for screening the substrate specificities of protein kinase. However, a solid phase assay using a peptide library-bound polymer resin or a peptide-arrayed glass chip is preferred for high throughput screening (HTS).
2010 Wiley Periodicals, Inc.
Similar articles
-
A high-throughput, nonisotopic, competitive binding assay for kinases using nonselective inhibitor probes (ED-NSIP).J Biomol Screen. 2002 Dec;7(6):507-14. doi: 10.1177/1087057102238624. J Biomol Screen. 2002. PMID: 14599348
-
Antibody-free peptide substrate screening of serine/threonine kinase (protein kinase A) with a biotinylated detection probe.Anal Biochem. 2011 Jun 1;413(1):30-5. doi: 10.1016/j.ab.2011.02.005. Epub 2011 Mar 3. Anal Biochem. 2011. PMID: 21310143
-
Development of high-throughput phosphorylation profiling method for identification of Ser/Thr kinase specificity.J Pept Sci. 2011 May;17(5):392-7. doi: 10.1002/psc.1312. Epub 2010 Nov 22. J Pept Sci. 2011. PMID: 21480437
-
Application of combinatorial library methods in cancer research and drug discovery.Anticancer Drug Des. 1997 Apr;12(3):145-67. Anticancer Drug Des. 1997. PMID: 9154108 Review.
-
Protein tyrosine kinases: structure, substrate specificity, and drug discovery.Biopolymers. 1998;47(3):197-223. doi: 10.1002/(SICI)1097-0282(1998)47:3<197::AID-BIP2>3.0.CO;2-H. Biopolymers. 1998. PMID: 9817025 Review.
Cited by
-
Targeted drug delivery for cancer therapy: the other side of antibodies.J Hematol Oncol. 2012 Nov 9;5:70. doi: 10.1186/1756-8722-5-70. J Hematol Oncol. 2012. PMID: 23140144 Free PMC article. Review.
-
Systematic analysis of phosphotyrosine antibodies recognizing single phosphorylated EPIYA-motifs in CagA of East Asian-type Helicobacter pylori strains.BMC Microbiol. 2016 Sep 2;16(1):201. doi: 10.1186/s12866-016-0820-6. BMC Microbiol. 2016. PMID: 27590005 Free PMC article.
-
Photocleavable peptide-conjugated magnetic beads for protein kinase assays by MALDI-TOF MS.Bioconjug Chem. 2010 Oct 20;21(10):1917-24. doi: 10.1021/bc1003058. Bioconjug Chem. 2010. PMID: 20860375 Free PMC article.
-
Enzyme Activity Assays for Protein Kinases: Strategies to Identify Active Substrates.Curr Drug Discov Technol. 2016;13(1):2-15. doi: 10.2174/1570163813666160115125930. Curr Drug Discov Technol. 2016. PMID: 26768716 Free PMC article. Review.
-
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.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
