Salmas Ramin Ekhteiari, Borysik Antoni James
Department of Chemistry, Britannia House, King's College London, SE1 1DB, London, UK.
Commun Biol. 2021 Feb 15;4(1):199. doi: 10.1038/s42003-021-01709-x.
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.
蛋白质免受氢氘交换（HDX）影响的程度为其折叠、动力学和相互作用提供了有价值的见解。通过质谱（MS）进行表征，HDX具有可忽略不计的质量限制以及出色的通量和灵敏度，但以分辨率为代价。由于根据蛋白水解消化程度，氨基酸以不同大小的组进行表征，所以无法准确确定或理解单个残基自然发生的交换机制。在此，我们报告了HDXmodeller，这是世界上首个用于高分辨率HDX-MS的在线网络服务器。HDXmodeller接受低分辨率HDX-MS输入数据，并返回为每个残基量化的高分辨率交换率。至关重要的是，HDXmodeller还会返回一组独特的统计数据，这些数据可以将交换率模型正确验证到99%的准确率。值得注意的是，这些统计数据无需任何关于单个交换率的先验知识即可得出，有助于增强用户信心，并具备评估不同数据优化策略的能力。
