Human adenovirus serotypes 3 and 5 bind to two different cellular receptors via the fiber head domain
Human adenovirus serotypes 3 and 5 bind to two different cellular receptors via the fiber head domain
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人腺病毒血清型 3 和 5 通过纤维头结构域与两种不同的细胞受体结合
DOI：
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发表时间：
1995
影响因子：
5.4
通讯作者：
Andalan Mcclelland
中科院分区：
文献类型：
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作者：
S. Stevenson;M. Rollence;Brian White;L. Weaver;Andalan Mcclelland
腺病毒纤维蛋白负责将病毒粒子附着到细胞表面受体上。介导结合的细胞受体的身份尚不清楚，但有证据表明两种不同的腺病毒受体与 C 组（5 型腺病毒 [Ad5]）和 B 组（Ad3）腺病毒相互作用。为了确定腺病毒受体特异性的决定因素，我们使用Ad5和Ad3的重组天然纤维多肽以及Ad5和Ad3的头结构域交换的嵌合纤维蛋白进行了一系列竞争结合实验。使用体外合成的放射性标记蛋白以及通过与杆状病毒表达的纤维蛋白的竞争分析来评估纤维与HeLa细胞受体的特异性结合。体外产生的纤维被发现为单体和三聚体，但只有组装的三聚体具有受体结合活性。竞争数据支持 Ad5 和 Ad3 与不同细胞受体相互作用的结论。使用纤维结合流式细胞术评估 Ad5 受体在几种细胞系上的分布。研究发现 HeLa 细胞表达高水平的受体，而 CHO 和人二倍体成纤维细胞则不然。含有 Ad5 纤维头结构域的嵌合纤维可阻断 Ad5 纤维的结合，但不能阻断 Ad3 纤维的结合。类似地，含有 Ad3 纤维头的嵌合纤维阻断了标记的 Ad3 纤维的结合，但不能阻断 Ad5 纤维的结合。此外，分离的 Ad3 纤维头结构域与标记的 Ad3 纤维有效竞争与 HeLa 细胞受体的结合。这些结果证明受体结合的决定因素位于纤维的头域中，并且分离的头域能够三聚化并与细胞受体结合。我们的结果还表明，通过操纵头域中包含的序列可以改变纤维蛋白的受体特异性。用新配体修饰或替换纤维头结构域可以允许具有新受体特异性的腺病毒载体，这可用于设计体内靶向基因递送。
The adenovirus fiber protein is responsible for attachment of the virion to cell surface receptors. The identity of the cellular receptor which mediates binding is unknown, although there is evidence suggesting that two distinct adenovirus receptors interact with the group C (adenovirus type 5 [Ad5]) and the group B (Ad3) adenoviruses. In order to define the determinants of adenovirus receptor specificity, we have carried out a series of competition binding experiments using recombinant native fiber polypeptides from Ad5 and Ad3 and chimeric fiber proteins in which the head domains of Ad5 and Ad3 were exchanged. Specific binding of fiber to HeLa cell receptors was assessed with radiolabeled protein synthesized in vitro, and by competition analysis with baculovirus-expressed fiber protein. Fiber produced in vitro was found as both monomer and trimer, but only the assembled trimers had receptor binding activity. Competition data support the conclusion that Ad5 and Ad3 interact with different cellular receptors. The Ad5 receptor distribution on several cell lines was assessed with a fiber binding flow cytometric assay. HeLa cells were found to express high levels of receptor, while CHO and human diploid fibroblasts did not. A chimeric fiber containing the Ad5 fiber head domain blocked the binding of Ad5 fiber but not Ad3 fiber. Similarly, a chimeric fiber containing the Ad3 fiber head blocked the binding of labeled Ad3 fiber but not Ad5 fiber. In addition, the isolated Ad3 fiber head domain competed effectively with labeled Ad3 fiber for binding to HeLa cell receptors. These results demonstrate that the determinants of receptor binding are located in the head domain of the fiber and that the isolated head domain is capable of trimerization and binding to cellular receptors. Our results also show that it is possible to change the receptor specificity of the fiber protein by manipulation of sequences contained in the head domain. Modification or replacement of the fiber head domain with novel ligands may permit adenovirus vectors with new receptor specificities which could be useful for targeted gene delivery in vivo to be engineered.
腺病毒纤维蛋白的氨基末端编码核定位信号。
影响因子：
3.7
作者：
Hong,JS;Engler,JA
通讯作者：
Engler,JA
影响因子：
56.9
作者：
KASAHARA, N;DOZY, AM;KAN, YW
通讯作者：
KAN, YW