High-throughput screening for protein tyrosine phosphatase activity modulators.

摘要: Reversible phosphorylation of proteins, principally on serine, threonine, or tyrosine residues, is central to the regulation most aspects eukaryotic cell function. Dysregulation protein kinases and phosphatases linked numerous human diseases. Consequently, many efforts have been made target these enzymes with small molecules in order develop new therapeutic agents. While kinase inhibitors successfully brought market, development specific phosphatase still its infancy. The largest diverse superfamily humans comprised by phosphatases, a group over 100 enzymes. Here, we describe high-throughput screening methods search for activity modulators. We illustrate implementation relatively simple assays, using generic absorbance- fluorescence-based substrates, 384- 1536-well microtiter plates. discuss steps optimize HTS assay quality performance, several PTP basis previously performed successful campaigns. Finally, how confirm, follow up, prioritize hit compounds, point out number common pitfalls that are encountered this process.