Identification of Macrolide Antibiotic-binding Human_p8 Protein

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.

The role of antibiotics and antibiotic resistance in nature

“…Tetracyclines, macrolides and ketolides display potent anti‐inflammatory activities (Tamaoki et al ., 2004; Weinberg, 2005; Del Rosso, 2007; Webster and Del Rosso, 2007; Leiva et al ., 2008a,b) while rifampin exerts the opposite effect (Yuhas et al ., 2008). The macrolide antibiotic‐binding human p8 protein was recently cloned and identified using the phage display library approach (Morimura et al ., 2008). This is a nuclear DNA‐binding protein, which is strongly activated in response to several stresses, and, on the basis of functional similarity to HMG‐I/Y‐like proteins, it was suggested that p8 may be involved in transcription regulation (Encinar et al ., 2001; Hoffmeister et al ., 2002).…”

“…This is a nuclear DNA‐binding protein, which is strongly activated in response to several stresses, and, on the basis of functional similarity to HMG‐I/Y‐like proteins, it was suggested that p8 may be involved in transcription regulation (Encinar et al ., 2001; Hoffmeister et al ., 2002). Since clarithromycin, erythromycin and azithromycin inhibit the binding of recombinant p8 protein to double‐stranded DNA (Morimura et al ., 2008), the anti‐inflammatory effect of macrolides may be explained by the downregulation of transcription of genes involved in the pro‐inflammatory network. Interestingly, the same inhibitory effect was observed with a structurally unrelated antifungal antibiotic dechlorogriseofulvin (Morimura et al ., 2008) suggesting a potential overlap in recognition of structurally different antibiotic ligands by a single human molecular sensor.…”

“…Since clarithromycin, erythromycin and azithromycin inhibit the binding of recombinant p8 protein to double‐stranded DNA (Morimura et al ., 2008), the anti‐inflammatory effect of macrolides may be explained by the downregulation of transcription of genes involved in the pro‐inflammatory network. Interestingly, the same inhibitory effect was observed with a structurally unrelated antifungal antibiotic dechlorogriseofulvin (Morimura et al ., 2008) suggesting a potential overlap in recognition of structurally different antibiotic ligands by a single human molecular sensor.…”

Biotic acts of antibiotics

“…The macrolide antibiotic-binding human p8 protein has been cloned and identified using the phage display library approach (Morimura et al, 2008). This is a nuclear DNA-binding protein, which is strongly activated in response to several stresses, and, on the basis of functional similarity to HMG-I/Y-like proteins, it has been suggested that p8 may be involved in transcription regulation (Encinar et al, 2001; Hoffmeister et al, 2002).…”

“…In cardiac pathology, p8 is broadly involved in cellular events leading to cardiomyocyte hypertrophy and cardiac fibroblast MMPs production, both of which take place in heart failure (Goruppi et al, 2007). Since clarithromycin, erythromycin, and azithromycin inhibit the binding of recombinant p8 protein to double-stranded DNA (Morimura et al, 2008), the anti-inflammatory effect of macrolides discussed above may be explained, at least in part, by the down-regulation of transcription of genes involved in the proinflammatory network. Interestingly, the same inhibitory effect has been observed with a structurally unrelated antifungal antibiotic dechlorogriseofulvin (Morimura et al, 2008), suggesting a potential overlap in recognition of structurally different antibiotic ligands by a single human molecular target.…”