A Cysteine Zipper Stabilizes a Pre-Fusion F Glycoprotein Vaccine for Respiratory Syncytial Virus.
A Cysteine Zipper Stabilizes a Pre-Fusion F Glycoprotein Vaccine for Respiratory Syncytial Virus.
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DOI：
10.1371/journal.pone.0128779
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Kwong PD
中科院分区：
文献类型：
--
作者：
Stewart-Jones GB;Thomas PV;Chen M;Druz A;Joyce MG;Kong WP;Sastry M;Soto C;Yang Y;Zhang B;Chen L;Chuang GY;Georgiev IS;McLellan JS;Srivatsan S;Zhou T;Baxa U;Mascola JR;Graham BS;Kwong PD
重组亚单位疫苗应包含最少的非病原体基序，以减少潜在的脱靶反应性。我们最近开发了一种针对呼吸道合胞病毒（RSV）的疫苗抗原，它包含通过“DS - Cav1”突变以及来自T4噬菌体纤丝蛋白的附加C末端三聚化基序或“折叠域”而稳定在其融合前三聚体构象的融合（F）糖蛋白。在此，我们研究创建一个半胱氨酸拉链，以便在保持亲本DS - Cav1 + 折叠域抗原免疫原性的同时去除噬菌体折叠域。不含折叠域的构建体产生RSV F单体，并且从融合前F三聚体中酶促去除噬菌体折叠域导致它们解聚为单体。由于融合前RSV F胞外域的天然C末端包含一个病毒三聚体卷曲螺旋，我们探究了引入能够形成亚基间二硫键的半胱氨酸残基是否可能产生稳定的三聚体。结构建模表明引入的半胱氨酸形成二硫键“环”，每个环由卷曲螺旋的不同组向内的残基组成。三组环可置于天然RSV F卷曲螺旋内，并且通过复制卷曲螺旋的部分可添加额外的环。在小鼠中，一种无折叠域的4环稳定的RSV F三聚体引发了与亲本DS - Cav1 + 折叠域抗原相当的高水平中和活性。因此，基于结构对病毒卷曲螺旋进行改造以创建半胱氨酸拉链，使得可以从候选疫苗抗原中去除噬菌体三聚化基序。
Recombinant subunit vaccines should contain minimal non-pathogen motifs to reduce potential off-target reactivity. We recently developed a vaccine antigen against respiratory syncytial virus (RSV), which comprised the fusion (F) glycoprotein stabilized in its pre-fusion trimeric conformation by “DS-Cav1” mutations and by an appended C-terminal trimerization motif or “foldon” from T4-bacteriophage fibritin. Here we investigate the creation of a cysteine zipper to allow for the removal of the phage foldon, while maintaining the immunogenicity of the parent DS-Cav1+foldon antigen. Constructs without foldon yielded RSV F monomers, and enzymatic removal of the phage foldon from pre-fusion F trimers resulted in their dissociation into monomers. Because the native C terminus of the pre-fusion RSV F ectodomain encompasses a viral trimeric coiled-coil, we explored whether introduction of cysteine residues capable of forming inter-protomer disulfides might allow for stable trimers. Structural modeling indicated the introduced cysteines to form disulfide “rings”, with each ring comprising a different set of inward facing residues of the coiled-coil. Three sets of rings could be placed within the native RSV F coiled-coil, and additional rings could be added by duplicating portions of the coiled-coil. High levels of neutralizing activity in mice, equivalent to that of the parent DS-Cav1+foldon antigen, were elicited by a 4-ring stabilized RSV F trimer with no foldon. Structure-based alteration of a viral coiled-coil to create a cysteine zipper thus allows a phage trimerization motif to be removed from a candidate vaccine antigen.
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影响因子：
5.6
作者：
Grigoryan G;Degrado WF
通讯作者：
Degrado WF
影响因子：
3.7
作者：
Hotard AL;Shaikh FY;Lee S;Yan D;Teng MN;Plemper RK;Crowe JE Jr;Moore ML
通讯作者：
Moore ML
影响因子：
168.9
作者：
Lozano, Rafael;Naghavi, Mohsen;Murray, Christopher J. L.
通讯作者：
Murray, Christopher J. L.
影响因子：
5.4
作者：
Eggink, Dirk;Goff, Peter H.;Palese, Peter
通讯作者：
Palese, Peter
DOI：
10.1073/pnas.1115941109
发表时间：
2012-02-21
影响因子：
11.1
作者：
Magro, Margarita;Mas, Vicente;Palomo, Concepcion
通讯作者：
Palomo, Concepcion