| Autor: |
Zhang Lianwen, Ren Feifei, Li Jing, Ma Xiaofeng, Wang Peng |
| Jazyk: |
angličtina |
| Rok vydání: |
2009 |
| Předmět: |
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| Zdroj: |
Biological Procedures Online, Vol 11, Iss 1, Pp 170-183 (2009) |
| Druh dokumentu: |
article |
| ISSN: |
1480-9222 |
| Popis: |
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 (Em = 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. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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