Vers l’identification et la caractérisation des substrats des glutathion transférases de la classe phi de peuplier populus trichocarpa

Abstract : Glutathione transferases (GSTs) constitute a superfamily of enzymes found in most living organisms. These enzymes are notably involved in cellular detoxification processes as well as in the biosynthesis of secondary metabolites. An in silico analysis of Populus trichocarpa genome led to the identification of 8 genes encoding glutathione transferases from phi class. Recent work showed that these genes are expressed in both reproductive (female flowers and fruits) and vegetative organs (buds, leaves and petioles). These GSTs exhibit not only glutathionylation activities but also a peroxidase activity. The 3D structures of some isoforms have also been already solved in the absence of substrates in the H site. The identification of their natural substrates or ligands constitutes an essential step towards the understanding of their physiological roles within the cell. During this internship, secondary metabolites from male and female flowers as well as leaves and buds were extracted by maceration or by using a Soxhlet extractor. Potential substrates or ligands of the 8 GSTF isoforms, produced as recombinant proteins, were then searched from metaboliteenriched extracts previously prepared and from molecules of a chemical library by analyzing proteins thermostability as well as inhibition of esterase activity of these enzymes.
Document type :
Master thesis
File URL :
http://docnum.univ-lorraine.fr/prive/BUS_M_2015_PERROT_THOMAS.pdf
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https://hal.univ-lorraine.fr/hal-01769976
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Submitted on : Wednesday, April 18, 2018 - 3:02:03 PM
Last modification on : Thursday, April 19, 2018 - 1:13:26 AM

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Thomas Perrot. Vers l’identification et la caractérisation des substrats des glutathion transférases de la classe phi de peuplier populus trichocarpa. Sciences de l'environnement. 2015. ⟨hal-01769976⟩

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