Structure of a flavonoid glucosyltransferase reveals the basis for plant natural product modification.
Glycosylation is a key mechanism for orchestrating the bioactivity, metabolism and location of small molecules in living cells. In plants, a large multigene family of glycosyltransferases is involved in these processes, conjugating hormones, secondary metabolites, biotic and abiotic environmental toxins, to impact directly on cellular homeostasis. The red grape enzyme UDP-glucose:flavonoid 3-O-glycosyltransferase (VvGT1) is responsible for the formation of anthocyanins, the health-promoting compounds which, in planta, function as colourants determining flower and fruit colour and are precursors for the formation of pigmented polymers in red wine. We show that VvGT1 is active, in vitro, on a range of flavonoids. VvGT1 is somewhat promiscuous with respect to donor sugar specificity as dissected through full kinetics on a panel of nine sugar donors. The three-dimensional structure of VvGT1 has also been determined, both in its 'Michaelis' complex with a UDP-glucose-derived donor and the acceptor kaempferol and in complex with UDP and quercetin. These structures, in tandem with kinetic dissection of activity, provide the foundation for understanding the mechanism of these enzymes in small molecule homeostasis.
Department of Chemistry, University of York, York, UK.
Wendy
W
Offen
Carlos
C
Martinez-Fleites
Min
M
Yang
Eng
E
Kiat-Lim
Benjamin G
BG
Davis
Chris A
CA
Tarling
Christopher M
CM
Ford
Dianna J
DJ
Bowles
Gideon J
GJ
Davies
D018360:Crystallography, X-Ray; D005964:Glucosyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D044949:Kaempferols / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D011487:Protein Conformation; D011794:Quercetin / Q000378:metabolism*; D014530:Uridine Diphosphate / Q000378:metabolism*; D014532:Uridine Diphosphate Glucose / Q000378:metabolism*
D044949:Kaempferols; D014530:Uridine Diphosphate; C006552:kaempferol; D011794:Quercetin; D005964:Glucosyltransferases; C023249:flavonol 3-O-glucosyltransferase; D014532:Uridine Diphosphate Glucose