Gallo Carmela, Nuzzo Genoveffa, d'Ippolito Giuliana, Manzo Emiliano, Sardo Angela, Fontana Angelo
National Research Council of Italy, Institute of Biomolecular Chemistry, Pozzuoli, Naples, Italy.
National Research Council of Italy, Institute of Biomolecular Chemistry, Pozzuoli, Naples, Italy.
Methods Enzymol. 2018;605:101-138. doi: 10.1016/bs.mie.2018.03.003. Epub 2018 May 21.
Sterol sulfates are widely occurring molecules in marine organisms. Their importance has been so far underestimated although many of these compounds are crucial mediators of physiological and ecological functions in other organisms. Biosynthesis of sterol sulfates is controlled by cytosolic sulfotransferases (SULTs), a varied family of enzymes that catalyze the transfer of a sulfo residue (-SOH) from the universal donor 3'-phosphoadenosine-5'-phosphosulfate to the hydroxyl function at C-3 of the steroid skeleton. The absence of molecular tools has been the main impediment to the development of a biosynthetic study of this class of compounds in marine organisms. In fact, there is very limited information about these enzymes in marine environments. SULT activity has, however, been reported in several marine species, and, recently, the production of sterol sulfates has been linked to the control of growth in marine diatoms. In this chapter, we describe methods for the study of sterol sulfates in this lineage of marine microalgae. The main aim is to provide the tools useful to deal with the biosynthesis and regulation of these compounds and to circumvent the bottleneck of the lack of molecular information. The protocols have been designed for marine diatoms, but most of the procedures can be used for other marine organisms.
硫酸甾醇是海洋生物中广泛存在的分子。尽管其中许多化合物是其他生物生理和生态功能的关键介质，但它们的重要性至今仍被低估。硫酸甾醇的生物合成由胞质磺基转移酶（SULTs）控制，这是一类多样的酶，催化磺基残基（-SOH）从通用供体3'-磷酸腺苷-5'-磷酸硫酸转移到甾体骨架C-3位的羟基上。缺乏分子工具一直是开展海洋生物中这类化合物生物合成研究的主要障碍。事实上，关于海洋环境中这些酶的信息非常有限。然而，已有报道称几种海洋物种具有SULT活性，最近，硫酸甾醇的产生与海洋硅藻的生长控制有关。在本章中，我们描述了研究这类海洋微藻中硫酸甾醇的方法。主要目的是提供有助于处理这些化合物生物合成和调控的工具，并克服缺乏分子信息这一瓶颈。这些方案是针对海洋硅藻设计的，但大多数程序可用于其他海洋生物。
