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Observation of the time-course for peptidoglycan lipid intermediate II polymerization by Staphylococcus aureus monofunctional transglycosylase
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Braddick, Darren, Sandhu, Sandeep, Roper, David I., Chappell, M. J. (Michael J.) and Bugg, Tim (2014) Observation of the time-course for peptidoglycan lipid intermediate II polymerization by Staphylococcus aureus monofunctional transglycosylase. Microbiology, Volume 160 (Part 8). pp. 1628-1636. doi:10.1099/mic.0.079442-0 ISSN 1350-0872.
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Official URL: http://dx.doi.org/10.1099/mic.0.079442-0
Abstract
The polymerization of lipid intermediate II by the transglycosylase activity of penicillin-binding proteins (PBPs) represents an important target for antibacterial action, but limited methods are available for quantitative assay of this reaction, or screening potential inhibitors. A new labelling method for lipid II polymerization products using Sanger’s reagent (fluoro-2,4-dinitrobenzene), followed by gel permeation HPLC analysis, has permitted the observation of intermediate polymerization products for Staphylococcus aureus monofunctional transglycosylase MGT. Peak formation is inhibited by 6 µM ramoplanin or enduracidin. Characterization by mass spectrometry indicates the formation of tetrasaccharide and octasaccharide intermediates, but not a hexasaccharide intermediate, suggesting a dimerization of a lipid-linked tetrasaccharide. Numerical modelling of the time-course data supports a kinetic model involving addition to lipid-linked tetrasaccharide of either lipid II or lipid-linked tetrasaccharide. Observation of free octasaccharide suggests that hydrolysis of the undecaprenyl diphosphate lipid carrier occurs at this stage in peptidoglycan transglycosylation.
Item Type: | Journal Article | ||||||||||
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry Faculty of Science, Engineering and Medicine > Engineering > Engineering Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) |
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Journal or Publication Title: | Microbiology | ||||||||||
Publisher: | Society for General Microbiology | ||||||||||
ISSN: | 1350-0872 | ||||||||||
Official Date: | August 2014 | ||||||||||
Dates: |
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Volume: | Volume 160 | ||||||||||
Number: | Part 8 | ||||||||||
Page Range: | pp. 1628-1636 | ||||||||||
DOI: | 10.1099/mic.0.079442-0 | ||||||||||
Status: | Peer Reviewed | ||||||||||
Publication Status: | Published | ||||||||||
Access rights to Published version: | Restricted or Subscription Access |
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