Identification and characterization of glycoprotein gH of human herpesvirus-6.
Identification and characterization of glycoprotein gH of human herpesvirus-6.
复制标题
人疱疹病毒 6 糖蛋白 gH 的鉴定和表征。
DOI：
10.1006/viro.1993.1272
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发表时间：
1993
期刊：
影响因子：
3.7
通讯作者：
Chandran,B
中科院分区：
文献类型：
--
作者：
Qian,G;Wood,C;Chandran,B
对人疱疹病毒-6变异株GS （HHV-6A(GS)）的8-kbBamHI-G DNA片段进行序列分析，发现了一个与其他疱疹病毒糖蛋白gH同源的糖蛋白编码基因，该基因被命名为HHV-6 gH （Josephset al.， J. viroll .65, 5597）。采用聚合酶链反应扩增HHV-6A(GS) gH基因的开放阅读框（ORF），并以trpE融合蛋白在细菌中表达。仅用含有15 ~ 469个氨基酸的ORF构建稳定的融合蛋白，并用于兔抗血清的培养。在他们对gH基因的体外转录-翻译分析中，添加微粒体膜导致未加工前体加工成约108k的多肽产物。这些体外合成的前体和产物形式被兔抗gH融合蛋白抗体免疫沉淀。兔抗gH抗体中和了HHV-6变体A株GS和变体B株Z-29的感染性，免疫沉淀反应鉴定出病毒粒子包膜相关的102k多肽是HHV-6变体A和变体B的糖蛋白gH。除了102k糖蛋白外，（35S）蛋氨酸标记的HHV-6A（GS）感染细胞也一致地共沉淀出58k和164k的非糖基化多肽，其部分肽图不同。但不是来自HHV-6B（Z-29）感染的细胞。两株HHV-6变异组的糖蛋白gH分子量相似，且兔抗HHV-6A(GS) gH抗体能中和两组的感染性，提示HHV-6变异组的糖蛋白gH功能保守。
Sequence analysis of an 8-kbBamHI-G DNA fragment from human herpesvirus-6 variant A strain GS (HHV-6A(GS)) has identified a gene encoding a glycoprotein homologous to glycoprotein gH of other herpesviruses and this gene was designated as HHV-6 gH (Josephset al., J. Virol.65, 5597). The open reading frame (ORF) of HHV-6A(GS) gH gene was amplified by polymerase chain reaction and was expressed as a trpE fusion protein in bacteria. Stable fusion protein was made only with constructs containing amino acids 15 to 469 of the ORF and this fusion protein was used to raise rabbit antisera. In thein vitrotranscription-translation analysis of the gH gene, addition of microsomal membranes resulted in the processing of an unprocessed precursor to a polypeptide product of about 108 K. Thesein vitro-synthesized precursor and product forms were immunoprecipitated by the rabbit antibodies against the gH fusion protein. Rabbit anti-gH antibodies neutralized the infectivity of both HHV-6 variant A strain GS and variant B strain Z-29, and immunoprecipitation reactions identified a virion envelope-associated 102 K polypeptide as the glycoprotein gH of HHV-6 variants A and B. In addition to the 102 K glycoprotein, nonglycosylated polypeptides of about 58 and 164 K with different partial peptide maps were also consistently coimmunoprecipitated from (35S]methionine-labeled HHV-6A(GS)-infected cells, but not from HHV-6B(Z-29)-infected cells. The similarity in the molecular weights of glycoprotein gH among the two strains belonging to the two variant groups of HHV-6 and neutralization of infectivity of both groups by rabbit antibodies against HHV-6A(GS) gH suggests conserved functions of glycoprotein gH among the two variant groups of HHV-6.