Effects on sialic acid recognition of amino acid mutations in the carbohydrate-binding cleft of the rotavirus spike protein
Mark J Kraschnefski, Andrea Bugarcic, Fiona E Fleming, Xing Yu, Mark von Itzstein, Barbara S Coulson, Helen Blanchard
GLYCOBIOLOGY | OXFORD UNIV PRESS INC | Published : 2009
The rotavirus spike protein VP4 mediates attachment to host cells and subsequent membrane penetration. The VP8(*) domain of VP4 forms the spike tips and is proposed to recognize host-cell surface glycans. For sialidase-sensitive rotaviruses such as rhesus (RRV), this recognition involves terminal sialic acids. We show here that the RRV VP8(*)(64-224) protein competes with RRV infection of host cells, demonstrating its relevance to infection. In addition, we observe that the amino acids revealed by X-ray crystallography to be in direct contact with the bound sialic acid derivative methyl alpha-D-N-acetylneuraminide, and that are highly conserved amongst sialidase-sensitive rotaviruses, are re..View full abstract
Awarded by National Health and Medical Research Council of Australia
HB, BSC, and MvI gratefully acknowledge the financial support of the Australian Research Council (ARC). HB thanks Dr. Jean-Luc Ferrer for assistance in operation of X-ray diffraction data collection equipment at beamline FIP-BM30A (ESRF). We thank Dr. Alex Szyczew and Dr. Thomas Haselhorst (Institute for Glycomics) for providing Neu5Ac alpha 2Me and training XY in STD-NMR, respectively. Professor Istvan Toth and Mr. Daniel Coles (University of Queensland) provided access to the MicroCalorimeter and helpful advice on ITC. BSC is a Senior Research Fellow (350253) of the National Health and Medical Research Council of Australia. MvI thanks the ARC for the award of an Australian Federation Fellowship.