title: Domain requirement of moenomycin binding to bifunctional transglycosylases and development of high-throughput discovery of antibiotics.
abstract: 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.
authors.affiliation: Genomics Research Center, Academia Sinica, Sec. 2, 128 Academia Road, Nangang, Taipei 115, Taiwan.
authors.forename: Ting-Jen Rachel
authors.initials: TJ
authors.lastname: Cheng
authors.forename: Ming-Ta
authors.initials: MT
authors.lastname: Sung
authors.forename: Hsin-Yu
authors.initials: HY
authors.lastname: Liao
authors.forename: Yi-Fan
authors.initials: YF
authors.lastname: Chang
authors.forename: Chia-Wei
authors.initials: CW
authors.lastname: Chen
authors.forename: Chia-Ying
authors.initials: CY
authors.lastname: Huang
authors.forename: Lien-Yang
authors.initials: LY
authors.lastname: Chou
authors.forename: Yen-Da
authors.initials: YD
authors.lastname: Wu
authors.forename: Yin-Hsuan
authors.initials: YH
authors.lastname: Chen
authors.forename: Yih-Shyun E
authors.initials: YS
authors.lastname: Cheng
authors.forename: Chi-Huey
authors.initials: CH
authors.lastname: Wong
authors.forename: Che
authors.initials: C
authors.lastname: Ma
authors.forename: Wei-Chieh
authors.initials: WC
authors.lastname: Cheng
mesh_terms: 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
chemical_list: D009844:Oligosaccharides; D046915:Penicillin-Binding Proteins; D016695:Glycosyltransferases; C419802:moenomycin