CHARACTERIZATION OF THE KNOB DOMAIN OF THE ADENOVIRUS TYPE-5 FIBER PROTEIN EXPRESSED IN ESCHERICHIA-COLI
CHARACTERIZATION OF THE KNOB DOMAIN OF THE ADENOVIRUS TYPE-5 FIBER PROTEIN EXPRESSED IN ESCHERICHIA-COLI
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DOI：
10.1128/jvi.68.8.5239-5246.1994
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发表时间：
1994-08-01
影响因子：
5.4
通讯作者：
GERARD, RD
中科院分区：
文献类型：
--
作者：
HENRY, LJ;XIA, D;GERARD, RD
腺病毒纤维蛋白用于将病毒附着在细胞表面的特定受体上。在结构上，这种蛋白质由一个细长的轴组成，它从病毒衣壳的顶点伸出来，终止于一个被称为旋钮的球形结构域。为了验证旋钮是与细胞受体相互作用的结构域，我们从5型腺病毒中克隆并表达了旋钮，并在大肠杆菌中进行了单次重复。该蛋白经常规层析纯化，并具有与腺病毒受体相互作用的功能特征。重组旋钮结构域以3 × 10(9) M（-1）的亲和力与每个HeLa细胞结合约4700个位点，阻断腺病毒对人细胞的感染。针对旋钮产生的抗体也能阻断病毒感染。通过凝胶过滤和蛋白质晶体的x射线衍射分析，表明该旋结由21 kda亚基的三聚体组成。结果证实三聚体旋钮是腺病毒受体的附着配体。
The adenovirus fiber protein is used for attachment of the virus to a specific receptor on the cell surface. Structurally, the protein consists of a long, thin shaft that protrudes from the vertex of the virus capsid and terminates in a globular domain termed the knob. To verify that the knob is the domain which interacts with the cellular receptor, we have cloned and expressed the knob from adenovirus type 5 together with a single repeat of the shaft in Escherichia coli. The protein was purified by conventional chromatography and functionally characterized for its interaction with the adenovirus receptor. The recombinant knob domain bound about 4,700 sites per HeLa cell with an affinity of 3 x 10(9) M(-1) and blocked adenovirus infection of human cells. Antibodies raised against the knob also blocked virus infection. By gel filtration and X-ray diffraction analysis of protein crystals, the knob was shown to consist of a homotrimer of 21-kDa subunits. The results confirm that the trimeric knob is the ligand for attachment to the adenovirus receptor.