interferENZY: A Web-Based Tool for Enzymatic Assay Validation and Standardized Kinetic Analysis.
data pollution
enzyme kinetics
linearization method
reproducibility
webserver
Journal
Journal of molecular biology
ISSN: 1089-8638
Titre abrégé: J Mol Biol
Pays: Netherlands
ID NLM: 2985088R
Informations de publication
Date de publication:
28 05 2021
28 05 2021
Historique:
received:
27
05
2020
revised:
28
07
2020
accepted:
31
07
2020
pubmed:
10
8
2020
medline:
25
8
2021
entrez:
10
8
2020
Statut:
ppublish
Résumé
Enzymatic assays are widely employed to characterize important allosteric and enzyme modulation effects. The high sensitivity of these assays can represent a serious problem if the occurrence of experimental errors surreptitiously affects the reliability of enzyme kinetics results. We have addressed this problem and found that hidden assay interferences can be unveiled by the graphical representation of progress curves in modified reaction coordinates. To render this analysis accessible to users across all levels of expertise, we have developed a webserver, interferENZY, that allows (i) an unprecedented tight quality control of experimental data, (ii) the automated identification of small and major assay interferences, and (iii) the estimation of bias-free kinetic parameters. By eliminating the subjectivity factor in kinetic data reporting, interferENZY will contribute to solving the "reproducibility crisis" that currently challenges experimental molecular biology. The interferENZY webserver is freely available (no login required) at https://interferenzy.i3s.up.pt.
Identifiants
pubmed: 32768452
pii: S0022-2836(20)30482-4
doi: 10.1016/j.jmb.2020.07.025
pii:
doi:
Substances chimiques
hen egg lysozyme
EC 3.2.1.-
Muramidase
EC 3.2.1.17
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Validation Study
Langues
eng
Sous-ensembles de citation
IM
Pagination
166613Informations de copyright
Copyright © 2020 Elsevier Ltd. All rights reserved.