The Receptor-Binding Domain of Influenza Virus Hemagglutinin Produced in Escherichia coli Folds into Its Native, Immunogenic Structure

被引:50
作者
DuBois, Rebecca M. [2 ]
Manuel Aguilar-Yanez, Jose [3 ]
Mendoza-Ochoa, Gonzalo I. [3 ]
Oropeza-Almazan, Yuriana [3 ]
Schultz-Cherry, Stacey [1 ]
Moises Alvarez, Mario [3 ]
White, Stephen W. [2 ]
Russell, Charles J. [1 ]
机构
[1] St Jude Childrens Res Hosp, Dept Infect Dis, Memphis, TN 38105 USA
[2] St Jude Childrens Res Hosp, Dept Biol Struct, Memphis, TN 38105 USA
[3] Ctr Biotecnol FEMSA, Monterrey 64849, Nuevo Leon, Mexico
基金
美国国家卫生研究院;
关键词
MEMBRANE-FUSION; PH; ANTIBODIES; GLYCOPROTEIN; CLEAVAGE; REGION; SITE;
D O I
10.1128/JVI.01412-10
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The hemagglutinin (HA) surface glycoprotein promotes influenza virus entry and is the key protective antigen in natural immunity and vaccines. The HA protein is a trimeric envelope glycoprotein consisting of a globular receptor-binding domain (HA-RBD) that is inserted into a membrane fusion-mediating stalk domain. Similar to other class I viral fusion proteins, the fusogenic stalk domain spontaneously refolds into its postfusion conformation when expressed in isolation, consistent with this domain being trapped in a metastable conformation. Using X-ray crystallography, we show that the influenza virus HA-RBD refolds spontaneously into its native, immunogenic structure even when expressed in an unglycosylated form in Escherichia coli. In the 2.10-angstrom structure of the HA-RBD, the receptor-binding pocket is intact and its conformational epitopes are preserved. Recombinant HA-RBD is immunogenic and protective in ferrets, and the protein also binds with specificity to sera from influenza virus-infected humans. Overall, the data provide a structural basis for the rapid production of influenza vaccines in E. coli. From an evolutionary standpoint, the ability of the HA-RBD to refold spontaneously into its native conformation suggests that influenza virus acquired this domain as an insertion into an ancestral membrane-fusion domain. The insertion of independently folding domains into fusogenic stalk domains may be a common feature of class I viral fusion proteins.
引用
收藏
页码:865 / 872
页数:8
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