Glycobiology Advance Access published online on September 14, 2006
Glycobiology, doi:10.1093/glycob/cwl051
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1 Institute for Glycomics, Griffith University (Gold Coast Campus), PMB 50 Gold Coast Mail Centre, Queensland 9726, Australia
* To whom correspondence should be addressed. 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 characterised by saturation transfer difference (STD) NMR spectroscopy. From this STD NMR data it is proposed that the VP8* core recognises an identical binding epitope in both Neu5Acalpha;2Me and the disaccharide Neu5Ac
Received July 17, 2006
Revised September 12, 2006
Accepted September 13, 2006
Article
STD NMR spectroscopy and molecular modelling investigation of the binding of N-acetylneuraminic acid derivatives to rhesus rotavirus VP8* core
Thomas Haselhorst 1, Helen Blanchard 1, Martin Frank 1, Mark J. Kraschnefski 1, Milton J. Kiefel 1, Alex J. Szyczew 1, Jeffery C. Dyason 1, Fiona E. Fleming 2, Gavan Holloway 2, Barbara S. Coulson 2, and Mark von Itzstein 1 *
2 Department of Microbiology and Immunology, The University of Melbourne, Victoria 3010, Australia
Mark von Itzstein, E-mail: m.vonitzstein{at}griffith.edu.au
Abstract 
(2,6)-s-Gal?
1Me. In the VP8*-disaccharide complex, the Neu5Ac moiety contributes 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 Neu5,9Ac2alpha;2Me in complex with 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 recognise the unsaturated Neu5Ac derivative Neu5Ac2en. Molecular modelling of the protein-Neu5Ac2en complex indicates that key interactions between the protein and the unsaturated Neu5Ac derivative when compared to Neu5Acalpha;2Me would not be sustained. Neu5Ac2Me, Neu5Ac(2,6)-s-Gal1Me, Neu5,9Ac22Me and Neu5Ac2en inhibited rotavirus infection of MA104 cells by 61%, 35%, 30% and 0% respectively at 10 mM concentration. NMR spectroscopic, molecular modelling and infectivity inhibition results are in excellent agreement and provide valuable information for the design of inhibitors of rotavirus infection.
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