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.
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Morimoto S, Takahashi Y, Watanabe Y, Omura S . Chemical modification of erythromycins. I. Synthesis and antibacterial activity of 6-O-methylerythromycins A. J Antibiot 37: 187–189 ( 1984)
Schunzen F, Zarivach R, Harms J, Bachan A, Tocilj A, Albrecht R, Yonath A, Franceschi F . Structural basis for the interaction of antibiotics with the peptidyl transferase centre in eubacteria. Nature 413: 814–821 ( 2001) and references therein.
Leung WK, Graham DY . Clarithromycin for Helicobacter pylori infection. Expert Opin Pharmacother 1: 507–514 ( 2000)
Jacks SS, Giguere S, Nguyen A . In vitro susceptibilities of Rhodococcus equi and other common equine pathogens to azithromycin, clarithromycin, and 20 other antimicrobials. Antimicrob Agents Chemother 47: 1742–1745 ( 2003)
Kurdowska A, Noble JM, Griffith DE . The effect of azithromycin and clarithromycin on ex vivo interleukin-8 (IL-8) release from whole blood and IL-8 production by human alveolar macrophages. J Antimicrob Chemother 47: 867–870 ( 2001)
Kikuchi T, Hagiwara K, Honda Y, Gomi K, Kobayashi T, Takahashi H, Tokue Y, Watanabe A, Nukiwa T . Clarithromycin suppresses lipopolysaccharide-induced interleukin-8 production by human monocytes through AP-1 and NF-kappa B transcription factors. J Antimicrob Chemother 49: 745–755 ( 2002)
Sasaki M, Ito T, Kashima M, Fukui S, Izumiyama N, Watanabe A, Sano M, Fujiwara Y, Miura M . Erythromycin and clarithromycin modulation of growth factor-induced expression of heparanase mRNA on human lung cancer cells in vitro. Mediators Inflamm 10: 259–267 ( 2001)
Yatsunami J, Hayashi S . Fourteen-membered ring macrolides as anti-angiogenic compounds. Anticancer Res 21: 4253–4258 ( 2001)
Morris TC, Ranaghan L, Morrison J . Phase II trial of clarithromycin and pamidronate therapy in myeloma. Med Oncol 18: 79–84 ( 2001)
Morimura T, Noda N, Kato N, Watanabe T, Saitoh T, Yamazaki T, Takata K, Aoki S, Ohta K, Ohshige M, Sakaguchi K, Sugawara F . Identification of antibiotic clarithromycin binding peptide displayed by T7 phage particles. J Antibiot 59: 625–632 ( 2006)
Casas-Finet JR, Kumar A, Morris G, Wilson SH, Karpel RL . Spectroscopic studies of the structural domains of mammalian DNA beta-polymerase. J Biol Chem 266: 19618–19625 ( 1991)
Encinar JA, Mallo GV, Mizyrycki C, Giono L, Gonzalez-Ros JM, Rico M, Canepa E, Moreno S, Neira JL, Iovanna JL . Human p8 is a HMG-I/Y-like protein with DNA binding activity enhanced by phosphorylation. J Biol Chem 276: 2742–2751 ( 2001)
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Morimura, T., Hashiba, M., Kameda, H. et al. Identification of Macrolide Antibiotic-binding Human_p8 Protein. J Antibiot 61, 291–296 (2008). https://doi.org/10.1038/ja.2008.41
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DOI: https://doi.org/10.1038/ja.2008.41