Skehel J J, Wiley D C
National Institute for Medical Research, London NW7 1AA, England.
Annu Rev Biochem. 2000;69:531-69. doi: 10.1146/annurev.biochem.69.1.531.
Hemagglutinin (HA) is the receptor-binding and membrane fusion glycoprotein of influenza virus and the target for infectivity-neutralizing antibodies. The structures of three conformations of the ectodomain of the 1968 Hong Kong influenza virus HA have been determined by X-ray crystallography: the single-chain precursor, HA0; the metastable neutral-pH conformation found on virus, and the fusion pH-induced conformation. These structures provide a framework for designing and interpreting the results of experiments on the activity of HA in receptor binding, the generation of emerging and reemerging epidemics, and membrane fusion during viral entry. Structures of HA in complex with sialic acid receptor analogs, together with binding experiments, provide details of these low-affinity interactions in terms of the sialic acid substituents recognized and the HA residues involved in recognition. Neutralizing antibody-binding sites surround the receptor-binding pocket on the membrane-distal surface of HA, and the structures of the complexes between neutralizing monoclonal Fabs and HA indicate possible neutralization mechanisms. Cleavage of the biosynthetic precursor HA0 at a prominent loop in its structure primes HA for subsequent activation of membrane fusion at endosomal pH (Figure 1). Priming involves insertion of the fusion peptide into a charged pocket in the precursor; activation requires its extrusion towards the fusion target membrane, as the N terminus of a newly formed trimeric coiled coil, and repositioning of the C-terminal membrane anchor near the fusion peptide at the same end of a rod-shaped molecule. Comparison of this new HA conformation, which has been formed for membrane fusion, with the structures determined for other virus fusion glycoproteins suggests that these molecules are all in the fusion-activated conformation and that the juxtaposition of the membrane anchor and fusion peptide, a recurring feature, is involved in the fusion mechanism. Extension of these comparisons to the soluble N-ethyl-maleimide-sensitive factor attachment protein receptor (SNARE) protein complex of vesicle fusion allows a similar conclusion.
血凝素(HA)是流感病毒的受体结合和膜融合糖蛋白,也是感染性中和抗体的作用靶点。通过X射线晶体学已确定了1968年香港流感病毒HA胞外域三种构象的结构:单链前体HA0;在病毒上发现的亚稳态中性pH构象,以及融合pH诱导的构象。这些结构为设计和解释关于HA在受体结合中的活性、新出现和再次出现的流行病的发生以及病毒进入过程中的膜融合等实验结果提供了框架。HA与唾液酸受体类似物复合物的结构,连同结合实验,从所识别的唾液酸取代基和参与识别的HA残基方面提供了这些低亲和力相互作用的细节。中和抗体结合位点围绕着HA膜远端表面上的受体结合口袋,中和性单克隆Fab与HA之间复合物的结构表明了可能的中和机制。生物合成前体HA0在其结构中的一个突出环处的切割使HA为随后在内体pH下激活膜融合做好准备(图1)。引发涉及将融合肽插入前体中的一个带电口袋;激活需要将其作为新形成的三聚体卷曲螺旋的N末端朝着融合靶膜挤出,并将C末端膜锚定重新定位到杆状分子同一端靠近融合肽的位置。将这种已形成用于膜融合的新HA构象与为其他病毒融合糖蛋白确定的结构进行比较表明,这些分子都处于融合激活构象,并且膜锚定和融合肽的并列(一个反复出现的特征)参与了融合机制。将这些比较扩展到囊泡融合的可溶性N - 乙基 - 马来酰亚胺敏感因子附着蛋白受体(SNARE)蛋白复合物可得出类似结论。