Probing a CMP-Kdn synthetase by 1H, 31P, and STD NMR spectroscopy.

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Title Probing a CMP-Kdn synthetase by 1H, 31P, and STD NMR spectroscopy.
Author Haselhorst, Thomas Erwin; Muenster-Kuehnel, Anja K.; Stolz, Anita; Oschiles, Melanie; Tiralongo, Giuseppe; Kitajima, Ken; Gerardy-Schahn, Rita; von Itzstein, Mark
Journal Name Biochemical and Biophysical Research Communications
Year Published 2005
Place of publication Amsterdam
Publisher Elsevier
Abstract 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.
Peer Reviewed Yes
Published Yes
Publisher URI http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/description#description
Alternative URI http://dx.doi.org/10.1016/j.bbrc.2004.12.040
Volume 327
Issue Number 2
Page from 565
Page to 570
ISSN 0006-291X
Date Accessioned 2006-02-10
Language en_AU
Research Centre Institute for Glycomics
Faculty Institute for Glycomics
Subject PRE2009-Medical Biochemistry: Carbohydrates
URI http://hdl.handle.net/10072/4183
Publication Type Journal Articles (Refereed Article)
Publication Type Code c1

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