title: A Modified Coupled Enzyme Method for O-linked GlcNAc Transferase Activity Assay.
abstract: In order to determine the activity of O-linked GlcNAc transferase (OGT), a modified coupled enzyme method was proposed. This method was based on the measurement of uridine 5'-(trihydrogen diphosphate) (UDP), a product generated in transglycosylation reaction. In the assay, UDP was coupled to the conversion of phosphoenolpyruvate to pyruvate using pyruvate kinase. Using a commercial pyruvate assay kit, the pyruvate was converted to a red terminal product, which could be photometrically measured at 570 nm or fluorometrically measured at 587 nm (E(m) = 535 nm) on a microplate reader. Kinetic study of a truncated recombinant mOGT and quantitative analysis of OGT in two biological samples indicated that this method was practical and competitive for quantitative analysis of OGT.
authors: Lianwen
authors: L
authors: Zhang
authors: Feifei
authors: F
authors: Ren
authors: Jing
authors: J
authors: Li
authors: Xiaofeng
authors: X
authors: Ma
authors: Peng
authors: P
authors: Wang