Contribution of cysteine residues in the extracellular domain of the F protein of human respiratory syncytial virus to its function.
Contribution of cysteine residues in the extracellular domain of the F protein of human respiratory syncytial virus to its function.
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DOI：
10.1186/1743-422x-3-34
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发表时间：
2006-05-24
期刊：
影响因子：
4.8
通讯作者：
Del Vecchio AM
中科院分区：
文献类型：
--
作者：
Day ND;Branigan PJ;Liu C;Gutshall LL;Luo J;Melero JA;Sarisky RT;Del Vecchio AM
所有已知的人呼吸道合胞病毒（HRSV）分离株的成熟F蛋白含有15个绝对保守的半胱氨酸（C）残基，这些残基在其他肺病毒以及副粘病毒的F蛋白中高度保守。为了探索HRSV F蛋白胞外结构域中的半胱氨酸对融合活性的贡献，将每个半胱氨酸改变为丝氨酸。半胱氨酸37、313、322、333、343、358、367、393、416和439的突变消除或大大降低了细胞表面表达，表明这些残基对于蛋白质正确折叠和转运至细胞表面至关重要。如所预期的，这些突变的融合活性大大降低或消除。在32°C、37°C或39.5°C下，半胱氨酸残基212、382和422的突变对细胞表面表达或融合活性几乎没有影响。F2亚基中C37和C69的突变分别消除或减少细胞表面表达75%。没有一个突变显示温度敏感表型。
The mature F protein of all known isolates of human respiratory syncytial virus (HRSV) contains fifteen absolutely conserved cysteine (C) residues that are highly conserved among the F proteins of other pneumoviruses as well as the paramyxoviruses. To explore the contribution of the cysteines in the extracellular domain to the fusion activity of HRSV F protein, each cysteine was changed to serine. Mutation of cysteines 37, 313, 322, 333, 343, 358, 367, 393, 416, and 439 abolished or greatly reduced cell surface expression suggesting these residues are critical for proper protein folding and transport to the cell surface. As expected, the fusion activity of these mutations was greatly reduced or abolished. Mutation of cysteine residues 212, 382, and 422 had little to no effect upon cell surface expression or fusion activity at 32°C, 37°C, or 39.5°C. Mutation of C37 and C69 in the F2 subunit either abolished or reduced cell surface expression by 75% respectively. None of the mutations displayed a temperature sensitive phenotype.