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.

酶促测定被广泛用于表征重要的变构和酶调节作用。如果实验错误的发生暗中影响了酶动力学结果的可靠性，则这些测定的高灵敏度可能代表一个严重的问题。我们已经解决了这个问题，并发现隐藏的分析干扰可以通过修改反应坐标中的进程曲线的图形表示来揭示。为了使所有专业知识水平的用户都可以访问此分析，我们开发了一个网络服务器 interferENZY，它允许 (i) 对实验数据进行前所未有的严格质量控制，(ii) 自动识别小的和主要的测定干扰，以及(iii) 无偏差动力学参数的估计。通过消除动力学数据报告中的主观因素，interferENZY 将有助于解决目前挑战实验分子生物学的“再现性危机”。interferENZY 网络服务器可在 https://interferenzy.i3s.up.pt 免费使用（无需登录）。