Substrate screening of protein kinases: detection methods and combinatorial peptide libraries.
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).
Kim
Mira
M
School of Chemical and Biological Engineering, Seoul National University, Seoul, 151-744 Korea.
Shin
Dong-Sik
DS
Kim
Jaehi
J
Lee
Yoon-Sik
YS
D000255:Adenosine Triphosphate / Q000737:chemistry*; D019151:Peptide Library; D010766:Phosphorylation; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics; D013379:Substrate Specificity
D019151:Peptide Library; D000255:Adenosine Triphosphate; D011494:Protein Kinases