The extent to which proteins are protected from hydrogen deuterium exchange (HDX) provides valuable insight into their folding, dynamics and interactions. Characterised by mass spectrometry (MS), HDX benefits from negligible mass restrictions and exceptional throughput and sensitivity but at the expense of resolution. Exchange mechanisms which naturally transpire for individual residues cannot be accurately located or understood because amino acids are characterised in differently sized groups depending on the extent of proteolytic digestion. Here we report HDXmodeller, the world’s first online webserver for high-resolution HDX-MS. HDXmodeller accepts low-resolution HDX-MS input data and returns high-resolution exchange rates quantified for each residue. Crucially, HDXmodeller also returns a set of unique statistics that can correctly validate exchange rate models to an accuracy of 99%. Remarkably, these statistics are derived without any prior knowledge of the individual exchange rates and facilitate unparallel user confidence and the capacity to evaluate different data optimisation strategies.

保护蛋白质免受氢氘交换 (HDX) 影响的程度为了解它们的折叠、动力学和相互作用提供了宝贵的见解。以质谱 (MS) 为特征，HDX 受益于可忽略的质量限制和出色的通量和灵敏度，但以牺牲分辨率为代价。无法准确定位或理解自然蒸发单个残基的交换机制，因为根据蛋白水解消化的程度，氨基酸以不同大小的组为特征。在这里，我们报告了 HDXmodeller，它是世界上第一个用于高分辨率 HDX-MS 的在线网络服务器。HDXmodeller 接受低分辨率 HDX-MS 输入数据并返回针对每个残基量化的高分辨率交换率。至关重要的是，HDXmodeller 还返回一组独特的统计数据，可以正确验证汇率模型，准确率为 99%。值得注意的是，这些统计数据是在没有任何个别汇率的先验知识的情况下得出的，并促进了无与伦比的用户信心和评估不同数据优化策略的能力。