interferENZY: A Web-Based Tool for Enzymatic Assay Validation and Standardized Kinetic Analysis

• Type: Article in International Scientific Journal
• Date: 2021

Authors

Maria Filipa Pinto (Author) Other; Antonio Baici (Author) Other; Pedro José Barbosa Pereira (Author) Other; Sandra Macedo Ribeiro (Author) Other; Annalisa Pastore (Author) Other; Fernando Rocha (Author) FEUP; Pedro M. Martins (Author) Other

Journal

Title: Journal of Molecular Biology

Serial No. 166613 Vol. 433 No. 11

Pages: 1-9

ISSN: 0022-2836

Publisher: Elsevier

Indexing

ISI Web of Knowledge - 1 Citation

Scopus - 1 Citation

Associated Projects

LEPABE - Financiamento Plurianual 2020-2023 da Unidade de I&D LEPABE - Laboratório de Engenharia de Processos, Ambiente, Biotecnologia e Energia

Other information

Authenticus ID: P-00S-MZR

DOI: 10.1016/j.jmb.2020.07.025

Abstract (EN): 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. © 2020 Elsevier Ltd

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 9