FluorescentTEM-1β-lactamasewithwild-typeactivityasa.pptVIP

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FluorescentTEM-1β-lactamasewithwild-typeactivityasa.ppt

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Fluorescent TEM-1 β-lactamase with wild-type activity as a rapid drug sensor for in?vitro drug screening by Wing-Lam Cheong, Ming-San Tsang, Pui-Kin So, Wai-Hong Chung, Yun-Chung Leung, and Pak-Ho Chan Biosci. Rep. Volume 34(5):e00136 October 1, 2014 ?2014 by Portland Press Ltd Structure of the TEM-1 β-lactamase The Val216 residue is shown in black. Wing-Lam Cheong et al. Biosci. Rep. 2014;34:e00136 ?2014 by Portland Press Ltd SDS/PAGE analysis of the wild type?and mutant forms of the TEM-1 β-lactamase Lane 1 (protein markers): rabbit muscle phosphorylase b (97400 Da), BSA (66200 Da), hen egg white ovalbumin (45 000 Da), bovine carbonic anhydrolase (31 000 Da), SBTI (soy-bean try... Wing-Lam Cheong et al. Biosci. Rep. 2014;34:e00136 ?2014 by Portland Press Ltd Chemical structures of the β-lactam antibiotics and β-lactamase inhibitor used in this study . Wing-Lam Cheong et al. Biosci. Rep. 2014;34:e00136 ?2014 by Portland Press Ltd Time-course fluorescence measurements of the labelled V216C mutant with different concentrations of β-lactam antibiotics and β-lactamase inhibitor (A) cefoxitin; (B) penicillin G; (C) ampicillin; (D) clavulanate. Wing-Lam Cheong et al. Biosci. Rep. 2014;34:e00136 ?2014 by Portland Press Ltd Time-course ESI–MS and fluorescence measurements on the binding of the labelled V216C mutant to cefoxitin The time-course fluorescence profile of the labelled V216C mutant and the [ES*]/[Etotal] values measured by ESI–MS at different time intervals are show... Wing-Lam Cheong et al. Biosci. Rep. 2014;34:e00136 ?2014 by Portland Press Ltd Molecular models of the fluorescein-labelled TEM-1 V216C mutant in the free-enzyme (E) and ES* states (A) Without penicillin G, the fluorescein molecule (orange) lies close to the active site of the enzyme. Wing-Lam Cheong et al. Biosci. Rep. 2014;34:e00136 ?2014 by Portland Press Ltd Time-course fluorescence measurements on the labelled V216C mutant in the presence of trypsin The fluorescence of the labelled V216C