Inhibition of CMP-sialic acid transport into golgi vesicles by nucleoside monophosphates

M. Chiaramonte, J. L. Koviach, C. Moore, V. V. Iyer, Carston R Wagner, R. L. Halcomb, W. Miller, P. Melançon, R. D. Kuchta

Research output: Contribution to journalArticle

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Abstract

We examined the interactions of nucleotides with the CMP-sialic acid transporter in order to better understand which features play a role in binding and to investigate the relationship between binding and subsequent transport. With respect to the sugar, the transporter requires a complete ribose ring for tight binding, and the 2′-ara hydrogen makes an important contact. The enzyme exhibits little specificity with respect to the 2′- and 3′-hydroxyls, as it tolerated substitutions ranging from fluorine to an azido group. In the base, the C4 amine and C2 carbonyl groups make important contacts, while the N3 nitrogen does not. However, adding a methyl group to N3 dramatically reduced binding, indicating that mass at this position sterically hinders binding. Adding a group at C5 had either no effect or slightly enhanced binding. To determine if the transporter recognizes these CMP analogues as substrates, we assayed them for their ability to trans stimulate CMP-sialic acid import. These data suggest that the enzyme transports a wide variety of NMPs, and the rate of transport is inversely proportional to the KI of the analogue. The importance of our findings for understanding the specificities of the different nucleotide-sugar tranlocators and the design of novel glycosylation inhibitors are discussed.

Original languageEnglish (US)
Pages (from-to)14260-14267
Number of pages8
JournalBiochemistry
Volume40
Issue number47
DOIs
StatePublished - Nov 27 2001

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Cytidine Monophosphate N-Acetylneuraminic Acid
Nucleosides
Sugars
Nucleotides
Glycosylation
Cytidine Monophosphate
Ribose
Fluorine
Enzymes
Hydroxyl Radical
Amines
Hydrogen
Substitution reactions
Nitrogen
Substrates

Cite this

Chiaramonte, M., Koviach, J. L., Moore, C., Iyer, V. V., Wagner, C. R., Halcomb, R. L., ... Kuchta, R. D. (2001). Inhibition of CMP-sialic acid transport into golgi vesicles by nucleoside monophosphates. Biochemistry, 40(47), 14260-14267. https://doi.org/10.1021/bi011262w

Inhibition of CMP-sialic acid transport into golgi vesicles by nucleoside monophosphates. / Chiaramonte, M.; Koviach, J. L.; Moore, C.; Iyer, V. V.; Wagner, Carston R; Halcomb, R. L.; Miller, W.; Melançon, P.; Kuchta, R. D.

In: Biochemistry, Vol. 40, No. 47, 27.11.2001, p. 14260-14267.

Research output: Contribution to journalArticle

Chiaramonte, M, Koviach, JL, Moore, C, Iyer, VV, Wagner, CR, Halcomb, RL, Miller, W, Melançon, P & Kuchta, RD 2001, 'Inhibition of CMP-sialic acid transport into golgi vesicles by nucleoside monophosphates', Biochemistry, vol. 40, no. 47, pp. 14260-14267. https://doi.org/10.1021/bi011262w
Chiaramonte M, Koviach JL, Moore C, Iyer VV, Wagner CR, Halcomb RL et al. Inhibition of CMP-sialic acid transport into golgi vesicles by nucleoside monophosphates. Biochemistry. 2001 Nov 27;40(47):14260-14267. https://doi.org/10.1021/bi011262w
Chiaramonte, M. ; Koviach, J. L. ; Moore, C. ; Iyer, V. V. ; Wagner, Carston R ; Halcomb, R. L. ; Miller, W. ; Melançon, P. ; Kuchta, R. D. / Inhibition of CMP-sialic acid transport into golgi vesicles by nucleoside monophosphates. In: Biochemistry. 2001 ; Vol. 40, No. 47. pp. 14260-14267.
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