title: A novel fluorogenic substrate for dinuclear Zn(II)-containing metallo-β-lactamases.
abstract: In an effort to prepare a fluorogenic substrate to be used in activity assays with metallo-β-lactamases, (6R,7R)-8-oxo-7-(2-oxo-2H-chromene-3-carboxamido)-3-((4-(2-oxo-2H-chromene-3-carboxamido)-phenylthio)methyl)-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (CA) was synthesized and characterized. CA exhibited a fluorescence quantum yield (φ) of 0.0059, two fluorescence lifetimes of 3.63×10(-10) and 5.38×10(-9)s, and fluorescence intensity that is concentration-dependent. Steady-state kinetic assays revealed that CA is a substrate for metallo-β-lactamases (MβLs) L1 and CcrA, exhibiting Km and kcat values of 18μM and 5s(-1) and 11μM and 17s(-1), respectively.
authors: Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, College of Chemistry and Materials Science, Northwest University, Xi'an 710069, PR China.
authors: Yi-Lin
authors: YL
authors: Zhang
authors: Yin-Lin
authors: YL
authors: Zhang
authors: Jian-Min
authors: JM
authors: Xiao
authors: Ji-Li
authors: JL
authors: Feng
authors: Ke-Wu
authors: KW
authors: Yang
authors: Lei
authors: L
authors: Feng
authors: Li-Sheng
authors: LS
authors: Zhou
authors: Michael W
authors: MW
authors: Crowder
mesh_terms: D053961:Azabicyclo Compounds / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D003374:Coumarins / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry*; D007700:Kinetics; D013056:Spectrophotometry, Ultraviolet; D013237:Stereoisomerism; D013379:Substrate Specificity; D015032:Zinc / Q000737:chemistry*; D001618:beta-Lactamases / Q000737:chemistry / Q000378:metabolism*
chemical_list: C583193:8-oxo-7-(2-oxo-2H-chromene-3-carboxamido)-3-((4-(2-oxo-2H-chromene-3-carboxamido)-phenylthio)methyl)-5-thia-1-azabicyclo(4.2.0)oct-2-ene-2-carboxylic acid; D053961:Azabicyclo Compounds; D003374:Coumarins; D005456:Fluorescent Dyes; D001618:beta-Lactamases; D015032:Zinc