A Screening Method for Potential Anti-pancreatic Cancer Candidate Compounds Based on Hsp90-Cdc37 Interaction Model in vitro
Hai Zhang, Sen Sun, Qingshan Chen, Guoqing Zhang, Zhenqing Zhang, Duxin Sun and Wenpeng ZhangAffiliation:
Department of Pharmaceutical Science, College of Pharmacy, University of Michigan, Ann Arbor 48109, USA.
AbstractThe Hsp90/Cdc37 (heat shock protein 90; cell division cycle 37) interaction is a promising target for anticancer candidate drug development. The purpose of this research is to establish a screening method for anti-cancer candidate compounds based on the Hsp90/Cdc37 interaction model in vitro by Split Renilla luciferase protein fragment-assisted complementation bioluminescence technology. The parameters influencing the protein binding in this model were optimized, including protein concentration, reaction time, and substrate concentration. The optimal condition was as follows: protein concentration 2µ M, the ratio of Hsp90 and Cdc37 1:1, reaction time 10 min, and the GLO® substrate (Promega, Madison, WI, USA) dilution 1:100. The screening model was further validated by some known compounds blocking the Hsp90/Cdc37 interactions. It was proved that this method is rapid and sensitive and could be used for screening of antipancreatic cancer candidate compounds.
Anti-cancer, Cdc37, a screening method, Hsp90, Hsp90/Cdc37 interaction, protein interaction model in vitro, SRLPFAC.
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