VP1 residues around the five-fold axis of enterovirus A71 mediate heparan sulfate interaction
VP1 residues around the five-fold axis of enterovirus A71 mediate heparan sulfate interaction
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DOI：
10.1016/j.virol.2016.11.009
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发表时间：
2017-01-15
期刊：
影响因子：
3.7
通讯作者：
Chan, Yoke Fun
中科院分区：
文献类型：
--
作者：
Tan, Chee Wah;Sam, I-Ching;Chan, Yoke Fun
肠道病毒A71（EV-A71）是一种嗜神经性肠道病毒，使用硫酸乙酰肝素作为附着受体。EV-A71-硫酸乙酰肝素相互作用的分子决定因素尚不清楚。通过计算机模拟肝素对接和VP 1中所有可能的赖氨酸残基的诱变，我们确定K162、K242和K244负责肝素相互作用和抑制。具有K242 A和K244 A的EV-A71突变体分别快速获得补偿突变T100 K或E98 A和Q145 R-T237 N，其恢复肝素结合表型。VP 1 -98和VP 1 -145均调节肝素结合。用VP 1-E98-E145完全消除肝素结合表型，但通过E98 K或E145 Q置换恢复。在细胞培养适应期间，EV-A71迅速获得K98或Q/G145以恢复肝素结合表型。结合下一代测序分析，我们的结果表明EV-A71在体外肝素适应过程中通过调节五重轴上的带正电荷残基具有高遗传可塑性。我们的发现对EV-A71疫苗生产、进化研究和发病机制产生了影响。
Enterovirus A71 (EV-A71) is a neurotropic enterovirus that uses heparan sulfate as an attachment receptor. The molecular determinants of EV-A71-heparan sulfate interaction are unknown. With In silico heparin docking and mutagenesis of all possible lysine residues in VP 1, we identified that K162, K242 and K244 are responsible for heparin interaction and inhibition. EV-A71 mutants with K242A and K244A rapidly acquired compensatory mutations, T100K or E98A, and Q145R-T237N respectively, which restored the heparin-binding phenotype. Both VP1-98 and VP1-145 modulates heparin binding. Heparin-binding phenotype was completely abolished with VP1-E98-E145, but was restored by an E98K or E145Q substitution. During cell culture adaptation, EV-A71 rapidly acquired K98 or Q/G145 to restore the heparin-binding phenotype. Together with next-generation sequencing analysis, our results implied that EV-A71 has high genetic plasticity by modulating positively charged residues at the five-fold axis during in vitro heparin adaptation. Our finding has impact on EV-A71 vaccine production, evolutionary studies and pathogenesis.