{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["June CM"],"funding":["NIAID NIH HHS"],"pagination":["70-4"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC4167909"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["463"],"pubmed_abstract":["By reacting fluorescein isothiocyanate with meropenem, we have prepared a carbapenem-based fluorescent ?-lactam. Fluorescein-meropenem binds both penicillin-binding proteins and ?-lactam sensors and undergoes a typical acylation reaction in the active site of these proteins. The probe binds the class D carbapenemase OXA-24/40 with close to the same affinity as meropenem and undergoes a complete catalytic hydrolysis reaction. The visible light excitation and strong emission of fluorescein render this molecule a useful structure-function probe through its application in sodium dodecyl sulfate-polyacrylamide gel electrophoresis assays as well as solution-based kinetic anisotropy assays. Its classification as a carbapenem ?-lactam and the position of its fluorescent modification render it a useful complement to other fluorescent ?-lactams, most notably Bocillin FL. In this study, we show the utility of fluorescein-meropenem by using it to detect mutants of OXA-24/40 that arrest at the acyl-intermediate state with carbapenem substrates but maintain catalytic competency with penicillin substrates."],"journal":["Analytical biochemistry"],"pubmed_title":["A fluorescent carbapenem for structure function studies of penicillin-binding proteins, ?-lactamases, and ?-lactam sensors."],"pmcid":["PMC4167909"],"funding_grant_id":["R01AI072219","R01 AI063517","R01AI063517","R15 AI082416","R01 AI072219","R15AI082416"],"pubmed_authors":["Ulberg LS","Vaughan RM","Leonard DA","June CM","Witucki LA","Bonomo RA"],"additional_accession":[]},"is_claimable":false,"name":"A fluorescent carbapenem for structure function studies of penicillin-binding proteins, ?-lactamases, and ?-lactam sensors.","description":"By reacting fluorescein isothiocyanate with meropenem, we have prepared a carbapenem-based fluorescent ?-lactam. Fluorescein-meropenem binds both penicillin-binding proteins and ?-lactam sensors and undergoes a typical acylation reaction in the active site of these proteins. The probe binds the class D carbapenemase OXA-24/40 with close to the same affinity as meropenem and undergoes a complete catalytic hydrolysis reaction. The visible light excitation and strong emission of fluorescein render this molecule a useful structure-function probe through its application in sodium dodecyl sulfate-polyacrylamide gel electrophoresis assays as well as solution-based kinetic anisotropy assays. Its classification as a carbapenem ?-lactam and the position of its fluorescent modification render it a useful complement to other fluorescent ?-lactams, most notably Bocillin FL. In this study, we show the utility of fluorescein-meropenem by using it to detect mutants of OXA-24/40 that arrest at the acyl-intermediate state with carbapenem substrates but maintain catalytic competency with penicillin substrates.","dates":{"release":"2014-01-01T00:00:00Z","publication":"2014 Oct","modification":"2020-10-31T08:51:21Z","creation":"2019-03-27T01:35:54Z"},"accession":"S-EPMC4167909","cross_references":{"pubmed":["25058926"],"doi":["10.1016/j.ab.2014.07.012"]}}