Screening of Paclitaxel-Binding Molecules from a Library of Random peptides Displayed on T7 Phage Particles Using paclitaxel-Photoimmobilized Resin

摘要

Paclitaxel (Taxol),an effective anticancer agent,is known to bind to tubulin and induce tubulin polymerization.Several other binding proteins of paclitaxel,such as Bcl-2,heat shock proteins,and NSC-1,have also been reported.Here,we describe a T7 phage-based display to screen for paclitaxel-binding molecules from a random peptide library using pachtaxel-photoimmobilized TentaGel resin.Specific phage particles that bind the paclitaxel-immobilized resin were obtained.Among them,two phage clones included the same consensus amino acid sequence (KACGRTRVTS).Analysis of the protein database using BLAST revealed that a portion of this sequence is conserved in the zinc finger domain of human NFX1.Binding affinity of paclitaxel against the partial recombinant protein of NFX1 (424aa-876aa) was confirmed by pull-down assays and surface plasmon resonance analyses.