Structure of a Flavonoid Glucosyltransferase Reveals the Basis for Plant Natural Product Modification

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2006 Feb 17

PMID 16482224

Citations 139

Authors

Wendy Offen

Carlos Martinez-Fleites

Min Yang

Eng Kiat-Lim

Benjamin G Davis

Chris A Tarling

Christopher M Ford

Dianna J Bowles

Gideon J Davies

Affiliations

Soon will be listed here.

Abstract

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.

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References

Hu Y, Chen L, Ha S, Gross B, Falcone B, Walker D . Crystal structure of the MurG:UDP-GlcNAc complex reveals common structural principles of a superfamily of glycosyltransferases. Proc Natl Acad Sci U S A. 2003; 100(3):845-9. PMC: 298689. DOI: 10.1073/pnas.0235749100. View

Terwilliger T, Berendzen J . Automated MAD and MIR structure solution. Acta Crystallogr D Biol Crystallogr. 1999; 55(Pt 4):849-61. PMC: 2746121. DOI: 10.1107/s0907444999000839. View

Lim E, Ashford D, Hou B, Jackson R, Bowles D . Arabidopsis glycosyltransferases as biocatalysts in fermentation for regioselective synthesis of diverse quercetin glucosides. Biotechnol Bioeng. 2004; 87(5):623-31. DOI: 10.1002/bit.20154. View

Jones P, Messner B, Nakajima J, Schaffner A, Saito K . UGT73C6 and UGT78D1, glycosyltransferases involved in flavonol glycoside biosynthesis in Arabidopsis thaliana. J Biol Chem. 2003; 278(45):43910-8. DOI: 10.1074/jbc.M303523200. View

Flint J, Taylor E, Yang M, Bolam D, Tailford L, Martinez-Fleites C . Structural dissection and high-throughput screening of mannosylglycerate synthase. Nat Struct Mol Biol. 2005; 12(7):608-14. DOI: 10.1038/nsmb950. View

Hu Y, Walker S . Remarkable structural similarities between diverse glycosyltransferases. Chem Biol. 2002; 9(12):1287-96. DOI: 10.1016/s1074-5521(02)00295-8. View

Tukey R, Strassburg C . Human UDP-glucuronosyltransferases: metabolism, expression, and disease. Annu Rev Pharmacol Toxicol. 2000; 40:581-616. DOI: 10.1146/annurev.pharmtox.40.1.581. View

Patenaude S, Seto N, Borisova S, Szpacenko A, Marcus S, Palcic M . The structural basis for specificity in human ABO(H) blood group biosynthesis. Nat Struct Biol. 2002; 9(9):685-90. DOI: 10.1038/nsb832. View

Esnouf R . An extensively modified version of MolScript that includes greatly enhanced coloring capabilities. J Mol Graph Model. 1997; 15(2):132-4, 112-3. DOI: 10.1016/S1093-3263(97)00021-1. View

10.

Davies G, Ducros V, Varrot A, Zechel D . Mapping the conformational itinerary of beta-glycosidases by X-ray crystallography. Biochem Soc Trans. 2003; 31(Pt 3):523-7. DOI: 10.1042/bst0310523. View

11.

Marcus S, Polakowski R, Seto N, Leinala E, Borisova S, Blancher A . A single point mutation reverses the donor specificity of human blood group B-synthesizing galactosyltransferase. J Biol Chem. 2003; 278(14):12403-5. DOI: 10.1074/jbc.M212002200. View

12.

Davies G, Gloster T, Henrissat B . Recent structural insights into the expanding world of carbohydrate-active enzymes. Curr Opin Struct Biol. 2005; 15(6):637-45. DOI: 10.1016/j.sbi.2005.10.008. View

13.

Yang M, Brazier M, Edwards R, Davis B . High-throughput mass-spectrometry monitoring for multisubstrate enzymes: determining the kinetic parameters and catalytic activities of glycosyltransferases. Chembiochem. 2005; 6(2):346-57. DOI: 10.1002/cbic.200400100. View

14.

Ford C, Boss P, Hoj P . Cloning and characterization of Vitis vinifera UDP-glucose:flavonoid 3-O-glucosyltransferase, a homologue of the enzyme encoded by the maize Bronze-1 locus that may primarily serve to glucosylate anthocyanidins in vivo. J Biol Chem. 1998; 273(15):9224-33. DOI: 10.1074/jbc.273.15.9224. View

15.

Yang M, Proctor M, Bolam D, Errey J, Field R, Gilbert H . Probing the breadth of macrolide glycosyltransferases: in vitro remodeling of a polyketide antibiotic creates active bacterial uptake and enhances potency. J Am Chem Soc. 2005; 127(26):9336-7. DOI: 10.1021/ja051482n. View

16.

Errey J, Mukhopadhyay B, Ravindranathan Kartha K, Field R . Flexible enzymatic and chemo-enzymatic approaches to a broad range of uridine-diphospho-sugars. Chem Commun (Camb). 2004; (23):2706-7. DOI: 10.1039/b410184g. View

17.

Bowles D, Isayenkova J, Lim E, Poppenberger B . Glycosyltransferases: managers of small molecules. Curr Opin Plant Biol. 2005; 8(3):254-63. DOI: 10.1016/j.pbi.2005.03.007. View

18.

Murshudov G, Vagin A, Dodson E . Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallogr D Biol Crystallogr. 1997; 53(Pt 3):240-55. DOI: 10.1107/S0907444996012255. View

19.

Ducros V, Zechel D, Murshudov G, Gilbert H, Szabo L, Stoll D . Substrate distortion by a beta-mannanase: snapshots of the Michaelis and covalent-intermediate complexes suggest a B(2,5) conformation for the transition state. Angew Chem Int Ed Engl. 2002; 41(15):2824-7. DOI: 10.1002/1521-3773(20020802)41:15<2824::AID-ANIE2824>3.0.CO;2-G. View

20.

Mulichak A, Losey H, WALSH C, Garavito R . Structure of the UDP-glucosyltransferase GtfB that modifies the heptapeptide aglycone in the biosynthesis of vancomycin group antibiotics. Structure. 2001; 9(7):547-57. DOI: 10.1016/s0969-2126(01)00616-5. View