Structural Determination of Uridine Diphosphate Glycosyltransferases Using X-Ray Crystallography

Methods Mol Biol. 2022:2396:227-241. doi: 10.1007/978-1-0716-1822-6_17.

Abstract

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.

Keywords: Crystallization; Crystallography; Diffraction; Molecular docking; Molecular replacement; Multiwavelength anomalous dispersion; Protein expression and purification; Uridine diphosphate glycosyltransferase.

MeSH terms

  • Crystallography, X-Ray*
  • Glycosylation
  • Glycosyltransferases / metabolism
  • Plants / metabolism
  • Uridine Diphosphate / metabolism

Substances

  • Uridine Diphosphate
  • Glycosyltransferases