Terbium(III) Luminescence-Based Assay for Esterase Activity.
Journal
Analytical chemistry
ISSN: 1520-6882
Titre abrégé: Anal Chem
Pays: United States
ID NLM: 0370536
Informations de publication
Date de publication:
02 07 2019
02 07 2019
Historique:
pubmed:
30
6
2019
medline:
20
9
2020
entrez:
29
6
2019
Statut:
ppublish
Résumé
Esterases catalyze the hydrolysis of esters to form a carboxylic acid and alcohol. These enzymes play a key role in both the detoxification of xenobiotic compounds and the metabolism of drugs and prodrugs. Numerous fluorogenic probes have been developed to monitor esterase activity. Most are based on an aromatic alcohol, and the others are based on an aromatic acid. These restrictions leave unexplored the specificity of esterases for aliphatic esters. Here, we report on the use of esters of thiopheneacetic acid coupled with the luminescence of terbium(III) as the basis for a continuous assay of esterase activity. This probe allows for a wide variation of the alcohol moiety and the detection of its hydrolysis at submicromolar concentrations. The assay verifies steady-state kinetic parameters for catalysis by pig liver esterase from either initial rates or the integration of progress curves, and its utility is evident with unpurified esterases in bacterial and human cell lysates.
Identifiants
pubmed: 31247727
doi: 10.1021/acs.analchem.9b01954
pmc: PMC7015665
mid: NIHMS1067146
doi:
Substances chimiques
Esters
0
Fluorescent Dyes
0
Terbium
06SSF7P179
Esterases
EC 3.1.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
8615-8621Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM044783
Pays : United States
Organisme : NIGMS NIH HHS
ID : R21 GM135780
Pays : United States
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