title: Trehalose-6-phosphate synthase from the cat flea Ctenocephalides felis and Drosophila melanogaster: gene identification, cloning, heterologous functional expression and identification of inhibitors by high throughput screening.
abstract: Trehalose phosphate synthase (EC 2.4.1.15; TPS) is the crucial enzyme for the biosynthesis of trehalose, the main haemolymph sugar of insects, and therefore a potential insecticidal molecular target. In this study, we report the functional heterologous expression of Drosophila melanogaster TPS, the gene identification, full length cDNA cloning and functional expression of cat flea (Ctenocephalides felis) TPS, and the Michaelis-Menten constants for their specific substrates glucose-6-phosphate and uridinediphosphate-glucose. A novel high throughput screening-compatible TPS assay and its use for the identification of the first potent insect TPS inhibitors from a large synthetic compound collection (>115 000 compounds) is described. One compound class that emerged in this screening, the 4-substituted 2,6-diamino-3,5-dicyano-4H-thiopyrans, was further investigated by analysing preliminary structure-activity relationships. Here, compounds were identified that show low µM to high nM half maximal inhibitory concentrations on insect TPS and that may serve as lead compounds for the development of insecticides with a novel mode of action.
authors: Intervet Innovation GmbH, Zur Propstei, 55270 Schwabenheim, Germany.
authors: C
authors: C
authors: Kern
authors: C
authors: C
authors: Wolf
authors: F
authors: F
authors: Bender
authors: M
authors: M
authors: Berger
authors: S
authors: S
authors: Noack
authors: S
authors: S
authors: Schmalz
authors: T
authors: T
authors: Ilg
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D058251:Ctenocephalides / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D019298:Glucose-6-Phosphate / Q000737:chemistry / Q000378:metabolism; D005964:Glucosyltransferases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007306:Insecticides / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D011714:Pyrans / Q000737:chemistry*; D054852:Small Molecule Libraries / Q000032:analysis / Q000737:chemistry; D013329:Structure-Activity Relationship; D014199:Trehalose / Q000235:genetics / Q000378:metabolism
chemical_list: D007306:Insecticides; D011714:Pyrans; D054852:Small Molecule Libraries; D019298:Glucose-6-Phosphate; D014199:Trehalose; D005964:Glucosyltransferases; C023180:trehalose-6-phosphate synthase