Establishment of in vitro Src tyrosine kinase inhibitors screening model

Abstract

A model system for rapid screening of Src tyrosine kinase inhibitors was established in vitro, and it is the foundation of screening tyrosine kinase activity inhibitors. The inclusion-bodies protein of GST-v-Src was bacterially expressed and collected, then, was actived by denature and renature. The activity of protein GST-v-Src and inhibitory ability toward Src tyrosine kinase was assayed by ELISA: Use a poly Glu∶Tyr (4∶1) as the substrate, and the fraction of phosphorylated substrate is visualized using a phosphotyrosine monoclonal antibody conjugated to HRP (HRP-PY20). This established model was high target specific, simple, convenient and high-throughput screening characteristics. By this method, six Chinese herbs in common use in cancer therapy were screened and three herbs containing abundant contents of Src tyrosine kinase inhibitory ability were picked out.

Keywords

• Src
• protein tyrosine kinase
• inhibitors
• screening
• Chinese herbs