Alderete Kasandra, Wang Xiaoqiang
Department of Biological Sciences and BioDiscovery Institute, University of North Texas, Denton, TX, USA.
Methods Mol Biol. 2022;2396:227-241. doi: 10.1007/978-1-0716-1822-6_17.
Uridine diphosphate glycosyltransferases (UGTs) are the key enzymes in glycosylation processes for decorating plant natural products with sugars. Crystallography, one of the powerful techniques for determining protein structures, was used as the main experimental technique and combined with biochemical methods to study the structure-function relationship and molecular mechanisms of UGTs. Crystal structures of plant UGTs have revealed their exquisite architectures and provided the structural basis for understanding their catalytic mechanism and substrate specificity. In this chapter, some protocols and experimental details of all key stages of protein structure determination are provided, and the structural insights on plant UGTs are also highlighted in combination of method description.
尿苷二磷酸糖基转移酶(UGTs)是用于给植物天然产物添加糖基修饰的糖基化过程中的关键酶。晶体学作为确定蛋白质结构的强大技术之一,被用作主要实验技术,并与生化方法相结合,以研究UGTs的结构-功能关系和分子机制。植物UGTs的晶体结构揭示了它们精妙的结构,并为理解其催化机制和底物特异性提供了结构基础。在本章中,提供了蛋白质结构测定所有关键阶段的一些方案和实验细节,并结合方法描述突出了对植物UGTs的结构见解。