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Updated: Jul 10, 2026
Biosensor-based High Throughput Biopanning and Bioinformatics Analysis Strategy for the Global Validation of Drug-protein Interactions
Published on: December 1, 2020
Screening of paclitaxel-binding molecules from a library of random peptides displayed on T7 phage particles using
Sota Aoki1, Kengo Morohashi, Takashi Sunoki
1Department of Applied Biological Science, Genome and Drug Research Center, and Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, Japan.
Abstract:
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 paclitaxel-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.
Insights
Researchers screened for paclitaxel-binding molecules using T7 phage display. They identified a peptide sequence conserved in human NFX1, confirming paclitaxel
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- Paclitaxel (Taxol) is a vital anticancer drug that interacts with tubulin.
- Previous studies identified various paclitaxel-binding proteins, including Bcl-2 and heat shock proteins.
Purpose of the Study:
- To identify novel paclitaxel-binding molecules using a phage display system.
- To investigate potential interactions between paclitaxel and human NFX1.
Main Methods:
- Utilized T7 phage display with paclitaxel-photoimmobilized TentaGel resin to screen a random peptide library.
- Identified specific phage clones exhibiting paclitaxel binding.
- Analyzed consensus peptide sequences using BLAST and confirmed binding affinity via pull-down assays and surface plasmon resonance (SPR).
Main Results:
- Successfully isolated phage particles that specifically bind to paclitaxel.
- Discovered a consensus amino acid sequence (KACGRTRVTS) in two phage clones.
- Identified a conserved motif within this sequence corresponding to the zinc finger domain of human NFX1.
- Confirmed paclitaxel binding to a partial recombinant NFX1 protein (424aa-876aa).
Conclusions:
- The T7 phage display system is effective for identifying paclitaxel-binding peptides.
- A novel interaction between paclitaxel and human NFX1 protein is suggested.
- Further research into NFX1 as a paclitaxel-binding protein may offer new therapeutic insights.
