Domain requirement of moenomycin binding to bifunctional transglycosylases and development of high-throughput discovery of antibiotics

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 105; no. 2; pp. 431 - 436
• Main Authors: Cheng, Ting-Jen Rachel, Sung, Ming-Ta, Liao, Hsin-Yu, Chang, Yi-Fan, Chen, Chia-Wei, Huang, Chia-Ying, Chou, Lien-Yang, Wu, Yen-Da, Chen, Yin-Hsuan, Cheng, Yih-Shyun E, Wong, Chi-Huey, Ma, Che, Cheng, Wei-Chieh
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 15.01.2008; National Acad Sciences
• Subjects: Anisotropy; anti-infective properties; Antibacterials; Antibiotics; Antimicrobials; bacteria; binding capacity; Binding sites; Biological Sciences; Cell growth; Cell Wall - metabolism; Cell walls; Chemistry, Pharmaceutical - methods; Correlation analysis; Drug Design; Enterococcus faecalis; Enterococcus faecalis - metabolism; Enzymes; Escherichia coli; fluorescence; Glycosyltransferases - chemistry; Inhibitory Concentration 50; Kinetics; Lipids; microbial growth; Models, Chemical; Molecules; NMR; Nuclear magnetic resonance; Oligosaccharides - chemistry; Oligosaccharides - pharmacology; Penicillin-Binding Proteins - chemistry; Physical Sciences; Protease inhibitors; Proteins; Spectrometry, Fluorescence - methods; Staphylococcus aureus; Staphylococcus aureus - metabolism; Staphylococcus infections; surface plasmon resonance; Surface Plasmon Resonance - methods; Technology, Pharmaceutical - methods
• ISSN: 0027-8424; 1091-6490; 1091-6490
• DOI: 10.1073/pnas.0710868105

Moenomycin inhibits bacterial growth by blocking the transglycosylase activity of class A penicillin-binding proteins (PBPs), which are key enzymes in bacterial cell wall synthesis. We compared the binding affinities of moenomycin A with various truncated PBPs by using surface plasmon resonance analysis and found that the transmembrane domain is important for moenomycin binding. Full-length class A PBPs from 16 bacterial species were produced, and their binding activities showed a correlation with the antimicrobial activity of moenomycin against Enterococcus faecalis and Staphylococcus aureus. On the basis of these findings, a fluorescence anisotropy-based high-throughput assay was developed and used successfully for identification of transglycosylase inhibitors.