Identification of macrolide antibiotic-binding Human_p8 protein
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Updated: Jul 3, 2026
Biosensor-based High Throughput Biopanning and Bioinformatics Analysis Strategy for the Global Validation of Drug-protein Interactions
Published on: December 1, 2020
Tetsuro Morimura1, Mio Hashiba, Hiroshi Kameda
1Genome and Drug Discovery Research Center, Department of Applied Biological Science, Tokyo University of Science, Noda, Chiba, Japan.
Abstract:
Clarithromycin is a macrolide antibiotic that is widely used in clinical medicine. Macrolide antibiotics such as clarithromycin specifically bind to the 50S subunit of the bacterial ribosome thereby interfering with protein biosynthesis. A selected peptide sequence from our former study, composed of 19 amino acids, which was isolated from a phage display library because of its ability to bind clarithromycin, displayed significant similarity to a portion of the human_p8 protein. The recombinant p8 protein binds to biotinylated-clarithromycin immobilized on a streptavidin-coated sensor chip and the dissociation constant was determined. The binding of recombinant p8 protein to double-stranded DNA was inhibited by biotinylated-clarithromycin, clarithromycin, erythromycin and azithromycin in gel mobility shift assay. Dechlorogriseofulvin, obtained from a natural product screening, also inhibited human p8 protein binding to DNA. This study illustrates the general utility of the phage display method in detecting protein-ligand interactions.
Insights
This study found that a human p8 protein fragment, identified using phage display, binds to the antibiotic clarithromycin. This interaction was further explored, revealing potential implications for drug discovery and understanding protein-ligand dynamics.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- Clarithromycin, a macrolide antibiotic, targets bacterial ribosomes and protein biosynthesis.
- Previous research identified a 19-amino acid peptide with clarithromycin-binding affinity via phage display.
- This peptide shares similarity with a segment of the human p8 protein.
Purpose of the Study:
- To investigate the interaction between the human p8 protein and clarithromycin.
- To characterize the binding affinity of the recombinant p8 protein to clarithromycin.
- To explore the influence of clarithromycin and related compounds on p8 protein-DNA binding.
Main Methods:
- Recombinant human p8 protein production and purification.
- Surface plasmon resonance (SPR) using streptavidin-coated sensor chips for binding studies.
- Gel mobility shift assays to assess inhibition of p8 protein-DNA interactions.
Main Results:
- The recombinant p8 protein demonstrated binding to biotinylated-clarithromycin, with a determined dissociation constant.
- Biotinylated-clarithromycin, clarithromycin, erythromycin, and azithromycin inhibited the binding of p8 protein to double-stranded DNA.
- Dechlorogriseofulvin also exhibited inhibitory effects on the p8 protein-DNA interaction.
Conclusions:
- The human p8 protein interacts with clarithromycin, suggesting a novel protein-ligand relationship.
- The phage display technique proved effective in identifying specific protein-ligand interactions.
- Findings highlight the potential for p8 protein as a target or modulator in drug-related studies.
