Biophysical measurement of the balance of influenza a hemagglutinin and neuraminidase activities.
Biophysical measurement of the balance of influenza a hemagglutinin and neuraminidase activities.
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DOI：
10.1074/jbc.m114.622308
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发表时间：
2015-03-06
期刊：
影响因子：
--
通讯作者：
McCauley JW
中科院分区：
文献类型：
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作者：
Benton DJ;Martin SR;Wharton SA;McCauley JW
背景资料：甲型流感病毒含有表面糖蛋白血凝素（HA）和神经氨酸酶（NA），分别负责受体结合和病毒释放。结果：使用生物层干涉测量法测量HA和NA对病毒与受体包被表面相互作用的贡献。结论：两种糖蛋白活性之间的平衡控制着病毒与细胞的相互作用，从而控制了病毒的传播性。意义：该技术可用于检查潜在的紧急病毒传播性的因素。甲型流感病毒与细胞表面的相互作用由表面糖蛋白血凝素（HA）和神经氨酸酶（NA）控制。这两种糖蛋白具有相反的活性：HA负责结合宿主受体（唾液酸）以允许感染，NA负责切割受体以促进病毒释放。几项研究表明，HA和NA活性的相容水平是病毒有效复制所需的。因此，这对于确定病毒的传播性具有很大的意义。这两种蛋白质在受体结合中的共同作用从未被直接测量过。我们展示了一种新的生物物理方法的基础上，生物层干涉测量这两种蛋白质的活动在真实的时间的平衡。该技术测量了在存在和不存在NA抑制剂的情况下，病毒与包被有人样受体类似物α 2，6-连接唾液酸或禽样受体类似物α 2，3-连接唾液酸的表面的结合和释放。还进行了生物层干涉测量以确定改变HA受体亲和力和NA茎长度对受体结合的影响。
Background: Influenza A viruses contain the surface glycoproteins hemagglutinin (HA) and neuraminidase (NA), responsible for receptor binding and virus release, respectively. Results: The contribution of HA and NA to virus interactions with receptor-coated surfaces was measured using bio-layer interferometry. Conclusion: The balance between the activities of the two glycoproteins controls virus-cell interactions, therefore transmissibility. Significance: This technique can be used to examine factors underlying emergent virus transmissibility. The interaction of influenza A viruses with the cell surface is controlled by the surface glycoproteins hemagglutinin (HA) and neuraminidase (NA). These two glycoproteins have opposing activities: HA is responsible for binding the host receptor (sialic acid) to allow infection, and NA is responsible for cleaving the receptor to facilitate virus release. Several studies have demonstrated that compatible levels of HA and NA activity are required for a virus to replicate efficiently. This is consequently of great interest for determining virus transmissibility. The concurrent role of these two proteins in receptor binding has never been directly measured. We demonstrate a novel biophysical approach based on bio-layer interferometry to measure the balance of the activities of these two proteins in real time. This technique measures virus binding to and release from a surface coated with either the human-like receptor analog α2,6-linked sialic acid or the avian-like receptor analog α2,3-linked sialic acid in both the presence and absence of NA inhibitors. Bio-layer interferometry measurements were also carried out to determine the effect of altering HA receptor affinity and NA stalk length on receptor binding.