title: Differential glycosylation produces heterogeneity in elevated esterases associated with insecticide resistance in the brown planthopper Nilaparvata lugens Stål.
abstract: The major insecticide resistance mechanism in the brown planthopper Nilaparvata lugens involves overproduction of esterases. Esterases purified from a resistant strain appeared as a ladder of bands on isoelectric focussing (IEF) gels from pI 4.7 to 5.0. Two-dimensional electrophoresis showed that isozymes ranged in size from 66 to 68 kDa with those of lower pI being apparently smaller. All isozymes detected by two-dimensional electrophoresis were glycosylated. N-glycosidase A reduced the number of isozymes on IEF to two, with increased pI and an increased molecular weight of 69 kDa. No O-linked glycans were detected. Deglycosylation had no effect on esterase activity, hence glycosylation is not involved in active site conformation. As N-glycosidase F completely deglycosylated the esterases, none of the glycans has an alpha1,3-bound core fucose. Reactivity with the lectins GNA, MAA and DSA, combined with differential cleavage of N-linked glycans with endoglycosidases F1 and F2, indicated that terminally linked mannose is present in high mannose and/or hybrid type glycans and that terminally linked sialic acid and galactose-beta(1-4)-N-acetylglucosamine are present in biantennary complexes. Neuraminidase treatment had the same effect on pI of isozymes as complete deglycosylation. Therefore, the majority of the heterogeneity of elevated esterases on IEF is due to differential attachment of sialic acid to glycans of the two proteins.
authors.affiliation: Cardiff School of Biosciences, University of Wales Cardiff, P.O. Box 915, Cardiff, UK. smallg@cardiff.ac.uk
authors.forename: G J
authors.initials: GJ
authors.lastname: Small
authors.forename: J
authors.initials: J
authors.lastname: Hemingway
mesh_terms: D000818:Animals; D004950:Esterases / Q000302:isolation & purification / Q000378:metabolism*; D006031:Glycosylation; D007313:Insecta / Q000201:enzymology*; D007305:Insecticide Resistance; D007526:Isoelectric Point; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D012794:Sialic Acids / Q000378:metabolism
chemical_list: D007527:Isoenzymes; D011134:Polysaccharides; D012794:Sialic Acids; D004950:Esterases