Virtual and Biophysical Screening Targeting the γ-Tubulin Complex – A New Target for the Inhibition of Microtubule Nucleation

摘要: Microtubules are the main constituents of mitotic spindles. They nucleated in large amounts during spindle assembly, from multiprotein complexes containing γ-tubulin and associated complex proteins (GCPs). With aim developing anti-cancer drugs targeting these nucleating complexes, we analyzed interface between GCP4 usually located a named γ-TuRC (γ-Tubulin Ring Complex). 10 ns molecular dynamics simulations were performed on heterodimers to obtain stable silico analyze residues involved persistent protein-protein contacts, responsible for stability complex. We demonstrated existence binding pocket at two upon formation. By combining virtual screening using fragment-based approach biophysical screening, found several small molecules that bind specifically this pocket. Sub-millimolar fragments have been experimentally characterized recombinant differential scanning fluorimetry (DSF) validation compounds as inhibitors. These results open new avenue drug development against microtubule-nucleating complexes.