Versatile Biosensor Deciphering Glycoenzymatic Activities

Session: 
PS2 Poster session 2 Even numbers
Code: 
P144
Location (hall): 
Foyer
Start/end time: 
Tuesday, July 2, 2019 - 15:45 to 17:15
Daniel
Marquez-Martin

Daniel Marquez-Martin1, Christine Saint-Pierre2, Aurelie Bouchet-Spinelli4, Olivier Lerouxel3, Didier Gasparutto2

1Grenoble Alpes University-Glyco@Alps, Grenoble, France, 2Atomic Energy and Alternative Energies Commission (CEA), Grenoble, France, 3Vegetal Macromolecules Research Centre (CERMAV), SAINT MARTIN D´HERES, France, 4Molecular SYstems and nanoMaterials for Energy and Health (SYMMES), Grenoble, France

To date, glycosyltransferases (GTs) are an essential family of enzymes poorly characterize both structurally and mechanistically which is being a major bottleneck in Glycoscience. They play a crucial role in living organisms catalysing the stereo- and regiospecific transfer of an activate donor sugar to an acceptor moiety to build up complex oligosaccharides onto the cell surface [1]. It requires the development of new methodologies able to elucidate and characterize the interactions between the glycosyl moiety and the enzyme. Surface plasmon resonance imaging (SPRi) has spread as a powerful analytical tool for deciphering many biomolecular processes exploring the kinetics and monitoring multiple interactions of biological and chemical substrates at real time and without any labels [2]. We aim at the development of a biosensor enable to rationalize the characterization of glycosyltransferases activities involved in the plant cell wall biosynthesis. To this aim, a fucosyltransferase from the plant Arabidosis thaliana (AtFUT1) (CAZY family GT37), was selected as model glycosyltransferase. This enzyme participates in the last step in the biosynthesis of xyloglucan (XG) transferring L-Fucose molecule from GDP-Fucose (donor) to a galactose sugar (acceptor) [3]. To that end, multiple chemical strategies to functionalize large moieties of xyloglucan to graft it on the gold surface have been developed [4] [5]. Depicting and monitoring the interactions between the AtFUT1 and the xyloglucan in terms of both specificity and activity represent a strong challenge. Complementary techniques such as isothermal titration calorimetry (iTC) may guide to streamline the comprehension and support the rationalization of the glycosylation process [6].

References: 
  1. Albesa-Jové, D; Guerin, M. Current Opinion in Structural Biology. 2016, 40, 23-32.
  2. Bosch, M; Sánchez, A.J; Rojas, F; Ojeda, C. Sensors. 2007, 7, 797-859.
  3. Cicéron, F; Rocha, J; Kousar, S; Hansen, S; Chazalet, V; Gillon, E; Breton, C; Lerouxel, O. Biochimie. 2016, 128-129, 183-192.
  4. Mallevre, F; Roget, A; Minon, T; Kervella, Y; Ropartz, D; Ralet, M.C; Canut, H; Livache, T. Bioconjugate Chem. 2013, 24, 1264-1269.
  5. Thakar, D; Migliorini, E; Coche-Guerente, L; Sadir, R; Lortat-Jacob, H; Boturyn, D; Renaudet, O; Labbe, P and Richter, R. Chem Commun. 2014, 50, 15148.
  6. Mazzei, L; Ciurli, S; Zambelli, B. Journal of Visualized Experiments. 2014, 86, 1-8.
Weight: 
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