Discovery of protein kinase phosphatase inhibitors.
Dynamic protein phosphorylation, a major cellular regulatory system, is tightly controlled by coordinating the reversible action of protein kinases and phosphatases. Recent evidence is consistent with sophisticated mechanisms that regulate both kinases and phosphatases in the cell. Dual specificity phosphatases, which act on phosphorylated serine, threonine, and tyrosine residues in proteins, are valid targets for drug discovery. Chemical complementation combines genetic manipulations with chemical biology and high-content multiparametric analyses and was developed as a screening approach to discover protein phosphatase inhibitors. Using a dual specificity mitogen-activated protein kinase phosphatase as an example, a detailed protocol, discussion of issues relating to data analysis, and high-throughput implementation of the chemical complementation approach to drug discovery is presented.
Vogt
Andreas
A
Department of Pharmacology, University of Pittsburgh Drug Discovery Institute, PA, USA.
Lazo
John S
JS
D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors*; D054638:Dual Specificity Phosphatase 1; D054642:Dual Specificity Phosphatase 6; D004791:Enzyme Inhibitors / Q000032:analysis* / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D017874:Immediate-Early Proteins / Q000037:antagonists & inhibitors*; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors*; D010766:Phosphorylation; D054645:Protein Phosphatase 1; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors*; D014162:Transfection
D018797:Cell Cycle Proteins; D004791:Enzyme Inhibitors; D017874:Immediate-Early Proteins; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1; C517019:DUSP1 protein, human; C517035:DUSP6 protein, human; D054638:Dual Specificity Phosphatase 1; D054642:Dual Specificity Phosphatase 6; D017027:Protein Tyrosine Phosphatases