Domain requirement of moenomycin binding to bifunctional transglycosylases and development of high-throughput discovery of antibiotics.
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.
Cheng
Ting-Jen Rachel
TJ
Genomics Research Center, Academia Sinica, Sec. 2, 128 Academia Road, Nangang, Taipei 115, Taiwan.
Sung
Ming-Ta
MT
Liao
Hsin-Yu
HY
Chang
Yi-Fan
YF
Chen
Chia-Wei
CW
Huang
Chia-Ying
CY
Chou
Lien-Yang
LY
Wu
Yen-Da
YD
Chen
Yin-Hsuan
YH
Cheng
Yih-Shyun E
YS
Wong
Chi-Huey
CH
Ma
Che
C
Cheng
Wei-Chieh
WC
D016880:Anisotropy; D002473:Cell Wall / Q000378:metabolism; D002626:Chemistry, Pharmaceutical / Q000379:methods; D015195:Drug Design; D013293:Enterococcus faecalis / Q000378:metabolism; D016695:Glycosyltransferases / Q000737:chemistry*; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008956:Models, Chemical; D009844:Oligosaccharides / Q000737:chemistry / Q000494:pharmacology*; D046915:Penicillin-Binding Proteins / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods; D013211:Staphylococcus aureus / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000379:methods; D013678:Technology, Pharmaceutical / Q000379:methods
D009844:Oligosaccharides; D046915:Penicillin-Binding Proteins; D016695:Glycosyltransferases; C419802:moenomycin