STD NMR spectroscopy and molecular modeling investigation of the binding of N-acetylneuraminic acid derivatives to rhesus rotavirus VP8* core

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Title STD NMR spectroscopy and molecular modeling investigation of the binding of N-acetylneuraminic acid derivatives to rhesus rotavirus VP8* core
Author Haselhorst, Thomas Erwin; Blanchard, Helen; Frank, Martin; Kraschnefski, Mark; Kiefel, Milton; Szyczew, Alexander Jeffery; Dyason, Jeffrey Clifford; Fleming, Fiona; Holloway, Gavan; Coulson, Barbara S.; von Itzstein, Mark
Journal Name Glycobiology
Year Published 2006
Place of publication Oxford, UK
Publisher Oxford University Press
Abstract The VP8* subunit of rotavirus spike protein VP4 contains a sialic acid (Sia)-binding domain important for host cell attachment and infection. In this study, the binding epitope of the N-acetylneuraminic acid (Neu5Ac) derivatives has been characterized by saturation transfer difference (STD) nuclear magnetic resonance (NMR) spectroscopy. From this STD NMR data, it is proposed that the VP8* core recognizes an identical binding epitope in both methyl -D-N-acetylneuraminide (Neu5Ac2Me) and the disaccharide methyl S-(-D-N-acetylneuraminosyl)-(26)-6-thio-ß-D-galactopyranoside (Neu5Ac-(2,6)-S-Galß1Me). In the VP8*–disaccharide complex, the Neu5Ac moiety contributes to the majority of interaction with the protein, whereas the galactose moiety is solvent-exposed. Molecular dynamics calculations of the VP8*–disaccharide complex indicated that the galactose moiety is unable to adopt a conformation that is in close proximity to the protein surface. STD NMR experiments with methyl 9-O-acetyl--D-N-acetylneuraminide (Neu5,9Ac22Me) in complex with rhesus rotavirus (RRV) VP8* revealed that both the N-acetamide and 9-O-acetate moieties are in close proximity to the Sia-binding domain, with the N-acetamide's methyl group being saturated to a larger extent, indicating a closer association with the protein. RRV VP8* does not appear to significantly recognize the unsaturated Neu5Ac derivative [2-deoxy-2,3-didehydro-D-N-acetylneuraminic acid (Neu5Ac2en)]. Molecular modeling of the protein–Neu5Ac2en complex indicates that key interactions between the protein and the unsaturated Neu5Ac derivative when compared with Neu5Ac2Me would not be sustained. Neu5Ac2Me, Neu5Ac-(2,6)-S-Galß1Me, Neu5,9Ac22Me, and Neu5Ac2en inhibited rotavirus infection of MA104 cells by 61%, 35%, 30%, and 0%, respectively, at 10 mM concentration. NMR spectroscopic, molecular modeling, and infectivity inhibition results are in excellent agreement and provide valuable information for the design of inhibitors of rotavirus infection.
Peer Reviewed Yes
Published Yes
Publisher URI http://glycob.oxfordjournals.org/
Alternative URI http://dx.doi.org/10.1093/glycob/cwl051
Copyright Statement This is a pre-copy-editing, author-produced PDF of an article accepted for publication in Glycobiology following peer review. The definitive publisher-authenticated version Glycobiology, Vol. 17(1), pp. 68-81 is available online at: http://dx.doi.org/10.1093/glycob/cwl051
Volume 17
Issue Number 1
Page from 68
Page to 81
ISSN 0959-6658
Date Accessioned 2007-02-23
Date Available 2009-09-08T08:04:15Z
Language en_AU
Research Centre Institute for Glycomics
Faculty Institute for Glycomics
Subject PRE2009-Biophysics; PRE2009-Medical Biochemistry: Carbohydrates; PRE2009-Structural Chemistry
URI http://hdl.handle.net/10072/13720
Publication Type Journal Articles (Refereed Article)
Publication Type Code c1

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