title: A Self-Immobilizing and Fluorogenic Probe for β-Lactamase Detection.
abstract: The spread of antibiotic resistance in pathogenic bacteria has become one of the major concerns to public health. Improved monitoring of drug resistance is of high importance for infectious disease control. One of the major mechanisms for bacteria to overcome treatment of antibiotics is the production of β-lactamases, which are enzymes that hydrolyze the β-lactam ring of the antibiotic. In this study, we have developed a self-immobilizing and fluorogenic probe for the detection of β-lactamase activity. This fluorogenic reagent, upon activation by β-lactamases, turns on a fluorescence signal and, more importantly, generates a covalent linkage to the target enzymes or the nearby proteins. The covalent labeling of enzymes was confirmed by SDS-PAGE analysis and MALDI-TOF mass spectrometry. The utility of this structurally simple probe was further confirmed by the fluorescent labeling of a range of β-lactamase-expressing bacteria.
authors: State Key Laboratory of Bioreactor Engineering, Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, 200237, P.R. China.
authors: Wuyu
authors: W
authors: Mao
authors: State Key Laboratory of Bioreactor Engineering, Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, 200237, P.R. China.
authors: Lingying
authors: L
authors: Xia
authors: State Key Laboratory of Bioreactor Engineering, Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, 200237, P.R. China.
authors: Yaqun
authors: Y
authors: Wang
authors: State Key Laboratory of Bioreactor Engineering, Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, 200237, P.R. China.
authors: Hexin
authors: http://orcid.org/0000-0003-1033-9211
authors: H
authors: Xie
mesh_terms: D002851:Chromatography, High Pressure Liquid; D003374:Coumarins / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004800:Enzymes, Immobilized / Q000032:analysis / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D006868:Hydrolysis; D008856:Microscopy, Fluorescence; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D001618:beta-Lactamases / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism
chemical_list: D003374:Coumarins; D004800:Enzymes, Immobilized; D005456:Fluorescent Dyes; C030123:coumarin; D001618:beta-Lactamases; C034413:beta-lactamase TEM-1
keywords: antibiotic resistance; enzymes; fluorescent probes; immobilization; β-lactamase