Versatile Biosensor Deciphering Glycoenzymatic Activities

PS2 Poster session 2 Even numbers
Location (hall): 
Start/end time: 
Tuesday, July 2, 2019 - 15:45 to 17:15

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].

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