Identification of macrolide antibiotic-binding Human_p8 protein.
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.
Morimura
Tetsuro
T
Genome and Drug Discovery Research Center, Department of Applied Biological Science, Tokyo University of Science, Noda, Chiba, Japan.
Hashiba
Mio
M
Kameda
Hiroshi
H
Takami
Mihoko
M
Takahama
Hirokazu
H
Ohshige
Masahiko
M
Sugawara
Fumio
F
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology; D051792:Basic Helix-Loop-Helix Proteins / Q000737:chemistry / Q000378:metabolism*; D001667:Binding, Competitive / Q000187:drug effects; D017291:Clarithromycin / Q000378:metabolism / Q000494:pharmacology; D004247:DNA / Q000378:metabolism; D030562:Databases, Protein; D024202:Electrophoretic Mobility Shift Assay; D006801:Humans; D018942:Macrolides / Q000378:metabolism* / Q000494:pharmacology; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000737:chemistry / Q000378:metabolism*; D010982:Platelet-Derived Growth Factor / Q000096:biosynthesis / Q000235:genetics; D011485:Protein Binding / Q000187:drug effects; D011994:Recombinant Proteins / Q000378:metabolism; D020349:Surface Plasmon Resonance
D000900:Anti-Bacterial Agents; D051792:Basic Helix-Loop-Helix Proteins; D017291:Clarithromycin; D004247:DNA; D018942:Macrolides; D009363:Neoplasm Proteins; D010982:Platelet-Derived Growth Factor; D011994:Recombinant Proteins; C491643:NUPR1 protein, human; C075278:platelet-derived growth factor A