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interferENZY: A Web-Based Tool for Enzymatic Assay Validation and Standardized Kinetic Analysis
Maria Filipa Pinto1, Antonio Baici2, Pedro José Barbosa Pereira3
1ICBAS-Instituto de Ciências Biomédicas Abel Salazar da Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal; LEPABE-Laboratório de Engenharia de Processos, Ambiente, Biotecnologia e Energia, Departamento de Engenharia Química, Faculdade de Engenharia da Universidade do Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal; i3S-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; IBMC-Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal.
Experimental errors can compromise enzyme kinetics. We developed interferENZY, a webserver that identifies assay interferences and provides reliable enzyme kinetic parameters, enhancing reproducibility in molecular biology.
Area of Science:
- Biochemistry and Molecular Biology
- Computational Biology and Cheminformatics
Background:
- Enzymatic assays are crucial for studying enzyme modulation but are sensitive to experimental errors.
- Unidentified assay interferences can lead to unreliable enzyme kinetics data and hinder reproducibility.
Purpose of the Study:
- To develop a method for detecting hidden assay interferences in enzymatic assays.
- To create a user-friendly webserver, interferENZY, for quality control and analysis of enzyme kinetics data.
Main Methods:
- Graphical analysis of progress curves in modified reaction coordinates to identify assay interferences.
- Development of the interferENZY webserver for automated data quality control and interference detection.
Main Results:
- The interferENZY webserver enables automated identification of assay interferences.
- It provides estimation of bias-free kinetic parameters, improving data reliability.
- The tool facilitates rigorous quality control of experimental data.
Conclusions:
- interferENZY offers a robust solution for detecting and correcting assay interferences in enzyme kinetics.
- This approach helps mitigate the reproducibility crisis in experimental molecular biology.
- The webserver democratizes access to reliable enzyme kinetic analysis.
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