کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1175583 | 961808 | 2006 | 6 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: In-gel activity staining of oxidized nicotinamide adenine dinucleotide kinase by blue native polyacrylamide gel electrophoresis In-gel activity staining of oxidized nicotinamide adenine dinucleotide kinase by blue native polyacrylamide gel electrophoresis](/preview/png/1175583.png)
Oxidized nicotinamide adenine dinucleotide (NAD+) kinase (NADK, E.C. 2.7.1.23) plays an instrumental role in cellular metabolism. Here we report on a blue native polyacrylamide gel electrophoretic technique that allows the facile detection of this enzyme. The product, oxidized nicotinamide adenine dinucleotide phosphate (NADP+), formed following the reaction of NADK with NAD+ and adenosine 5′-triphosphate was detected with the aid of glucose-6-phosphate dehydrogenase or NADP+–isocitrate dehydrogenase, iodonitrotetrazolium chloride, and phenazine methosulfate. The bands at the respective activity sites were excised and subjected to native and denaturing two-dimensional electrophoresis for the determination of protein levels. Hence this novel electrophoretic method allows the easy detection of NADK, a critical enzyme involved in pyridine homeostasis. Furthermore, this technique allowed the monitoring of the activity and expression of this kinase in various biological systems.
Journal: Analytical Biochemistry - Volume 359, Issue 2, 15 December 2006, Pages 210–215