JBIC Journal of Biological Inorganic Chemistry ( IF 2.7 ) Pub Date : 2020-09-14 , DOI: 10.1007/s00775-020-01818-8 Carlotta Zoppi 1 , Alessio Nocentini 2 , Claudiu T Supuran 2 , Alessandro Pratesi 3 , Luigi Messori 1
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Abstract
Native mass spectrometry is a potent technique to study and characterize biomacromolecules in their native state. Here, we have applied this method to explore the solution chemistry of human carbonic anhydrase I (hCA I) and its interactions with four different inhibitors, namely three sulfonamide inhibitors (AAZ, MZA, SLC-0111) and the dithiocarbamate derivative of morpholine (DTC). Through high-resolution ESI-Q-TOF measurements, the native state of hCA I and the binding of the above inhibitors were characterized in the molecular detail. Native mass spectrometry was also exploited to assess the direct competition in solution among the various inhibitors in relation to their affinity constants. Additional studies were conducted on the interaction of hCA I with the metallodrug auranofin, under various solution and instrumental conditions. Auranofin is a selective reagent for solvent-accessible free cysteine residues, and its reactivity was analyzed also in the presence of CA inhibitors. Overall, our investigation reveals that native mass spectrometry represents an excellent tool to characterize the solution behavior of carbonic anhydrase.
Graphic abstract
中文翻译：
人类碳酸酐酶I及其抑制剂复合物的天然质谱。
摘要
天然质谱法是研究和表征处于原始状态的生物大分子的有效技术。在这里，我们已应用此方法探索了人类碳酸酐酶I（hCA I）的溶液化学及其与四种不同抑制剂的相互作用，即三种磺酰胺抑制剂（AAZ，MZA，SLC-0111）和吗啉的二硫代氨基甲酸酯衍生物（DTC） ）。通过高分辨率的ESI-Q-TOF测量，在分子细节中表征了hCA I的天然状态和上述抑制剂的结合。还利用天然质谱法评估了各种抑制剂之间在亲和常数方面溶液的直接竞争。在各种溶液和仪器条件下，还进行了有关hCA I与金属药物金诺芬的相互作用的其他研究。金诺芬是一种用于溶剂可及的游离半胱氨酸残基的选择性试剂，并且在存在CA抑制剂的情况下也对其反应性进行了分析。总的来说，我们的研究表明，天然质谱是表征碳酸酐酶溶液行为的出色工具。
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