2P3I

Crystal structure of Rhesus Rotavirus VP8* at 295K

2P3I の概要

• エントリーDOI: 10.2210/pdb2p3i/pdb
• 関連するPDBエントリー: 2P3J 2P3K
• 分子名称: VP4, SULFATE ION, 2-O-methyl-5-N-acetyl-alpha-D-neuraminic acid, ... (4 entities in total)
• 機能のキーワード: beta-sandwich, viral protein
• 由来する生物種: Rhesus rotavirus
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 18575.42

構造登録者

Blanchard, H. (登録日: 2007-03-09, 公開日: 2008-03-11, 最終更新日: 2023-10-25)

主引用文献

Kraschnefski, M.J.,Bugarcic, A.,Fleming, F.E.,Yu, X.,von Itzstein, M.,Coulson, B.S.,Blanchard, H.
Effects on sialic acid recognition of amino acid mutations in the carbohydrate-binding cleft of the rotavirus spike protein
Glycobiology, 19:194-200, 2009

PubMed Abstract: 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 residues that are also important in interactions with host-cell carbohydrates. Residues Arg101 and Ser190 of the RRV VP8(*) carbohydrate-binding site were mutated to assess their importance for binding to the sialic acid derivative and their competition with RRV infection of host cells. The crystallographic structure of the Arg(101)Ala mutant crystallized in the presence of the sialic acid derivative was determined at 295 K to a resolution of 1.9 A. Our multidisciplinary study using X-ray crystallography, saturation transfer difference nuclear magnetic resonance spectroscopy, isothermal titration calorimetry, and competitive virus infectivity assays to investigate RRV wild-type and mutant VP8(*) proteins has provided the first evidence that the carbohydrate-binding cavity in RRV VP8(*) is used for host-cell recognition, and this interaction is not only with the sialic acid portion but also with other parts of the glycan structure.

PubMed: 18974199
DOI: 10.1093/glycob/cwn119

実験手法

X-RAY DIFFRACTION (1.75 Å)