Department of Medicinal Chemistry, Science for Life Laboratory, Uppsala University, Box 574, SE-75123 Uppsala, Sweden.
Department of Medicinal Chemistry, Science for Life Laboratory, Uppsala University, Box 574, SE-75123 Uppsala, Sweden.
Bioorg Med Chem. 2019 Mar 15;27(6):955-962. doi: 10.1016/j.bmc.2019.01.031. Epub 2019 Jan 30.
Sulfatases hydrolyze sulfated metabolites to their corresponding alcohols and are present in all domains of life. These enzymes have found major application in metabolic investigation of drugs, doping control analysis and recently in metabolomics. Interest in sulfatases has increased due to a link between metabolic processes involving sulfated metabolites and pathophysiological conditions in humans. Herein, we present the first comprehensive substrate specificity and kinetic analysis of the most commonly used arylsulfatase extracted from the snail Helix pomatia. In the past, this enzyme has been used in the form of a crude mixture of enzymes, however, recently we have purified this sulfatase for a new application in metabolomics-driven discovery of sulfated metabolites. To evaluate the substrate specificity of this promiscuous sulfatase, we have synthesized a series of new sulfated metabolites of diverse structure and employed a mass spectrometric assay for kinetic substrate hydrolysis evaluation. Our analysis of the purified enzyme revealed that the sulfatase has a strong preference for metabolites with a bi- or tricyclic aromatic scaffold and to a lesser extent for monocyclic aromatic phenols. This metabolite library and mass spectrometric method can be applied for the characterization of other sulfatases from humans and gut microbiota to investigate their involvement in disease development.
硫酸酯酶将硫酸化代谢物水解为相应的醇，存在于所有生命领域。这些酶在药物代谢研究、兴奋剂控制分析以及最近的代谢组学中得到了广泛应用。由于涉及硫酸化代谢物的代谢过程与人类病理生理状况之间存在联系，人们对硫酸酯酶的兴趣日益增加。本文首次对最常用的蜗牛 Helix pomatia 中提取的芳基硫酸酯酶进行了全面的底物特异性和动力学分析。过去，该酶一直以酶的粗混合物形式使用，但最近我们已经对该硫酸酯酶进行了纯化，以便在代谢组学驱动的硫酸化代谢物发现中进行新的应用。为了评估这种混杂硫酸酯酶的底物特异性，我们合成了一系列具有不同结构的新型硫酸化代谢物，并采用质谱法进行了动力学底物水解评估。我们对纯化酶的分析表明，该硫酸酯酶对具有双环或三环芳基支架的代谢物具有强烈的偏好，对单环芳基酚的偏好程度较低。该代谢物文库和质谱方法可用于鉴定来自人类和肠道微生物群的其他硫酸酯酶，以研究它们在疾病发展中的作用。
