کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
10771927 1050845 2005 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Probing a CMP-Kdn synthetase by 1H, 31P, and STD NMR spectroscopy
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Probing a CMP-Kdn synthetase by 1H, 31P, and STD NMR spectroscopy
چکیده انگلیسی
CMP-Kdn synthetase catalyses the reaction of sialic acids (Sia) and cytidine-5′-triphosphate (CTP) to the corresponding activated sugar nucleotide CMP-Sia and pyrophosphate PPi. STD NMR experiments of a recombinant nucleotide cytidine-5′-monophosphate-3-deoxy-d-glycero-d-galacto-nonulosonic acid synthetase (CMP-Kdn synthetase) were performed to map the binding epitope of the substrate CTP and the product CMP-Neu5Ac. The STD NMR analysis clearly shows that the anomeric proton of the ribose moiety of both investigated compounds is in close proximity to the protein surface and is likely to play a key role in the binding process. The relative rates of the enzyme reaction, derived from 1H NMR signal integrals, show that Kdn is activated at a rate 2.5 and 3.1 faster than Neu5Ac and Neu5Gc, respectively. Furthermore, proton-decoupled 31P NMR spectroscopy was successfully used to follow the enzyme reaction and clearly confirmed the appearance of CMP-Sia and the inorganic pyrophosphate by-product.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 327, Issue 2, 11 February 2005, Pages 565-570
نویسندگان
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