Rodriguez J R, Rodriguez D, Esteban M
Department of Biochemistry, State University of New York, Brooklyn 11203.
Virology. 1991 Apr;181(2):742-8. doi: 10.1016/0042-6822(91)90910-4.
To gain insights into the structure-function relationship of the envelope (env) glycoprotein of the human immunodeficiency virus type 1 (HIV-1) we have generated a vaccinia virus (VV) recombinant (VV-14kENV) that expresses a fusion protein (14k-env) consisting of the VV 14-kDa envelope protein (110 amino acids) fused at the C-terminus with HIV-1 env protein (816 amino acids). The 14k-env protein displayed unique structural properties in virus-infected cells. This protein was recognized by 14 kDa-specific antisera as well as HIV-1 env antisera. It was not cleaved during virus infection of cultured cells of various origins, it was stable, it was not released to the medium, and it was not incorporated into virions. Instead of a predicted 174-kDa protein, two proteins of about 110 and 100 kDa were observed. The size reduction of the fusion protein was due to limited glycosylation (110 kDa) and formation of unglycosylated protein (100 kDa). The 14k-env protein formed oligomeric structures and was exposed on the cell surface after virus infection. When mice were inoculated with the recombinant virus that expresses the 14K-env fusion protein, humoral immune response against gp160 was observed. Our findings suggest that 14k-env protein might display novel immunogenic properties.
为深入了解人类免疫缺陷病毒1型（HIV-1）包膜（env）糖蛋白的结构-功能关系，我们构建了一种痘苗病毒（VV）重组体（VV-14kENV），它表达一种融合蛋白（14k-env），该融合蛋白由VV的14 kDa包膜蛋白（110个氨基酸）在C端与HIV-1 env蛋白（816个氨基酸）融合而成。14k-env蛋白在病毒感染的细胞中表现出独特的结构特性。该蛋白可被14 kDa特异性抗血清以及HIV-1 env抗血清识别。在感染各种来源的培养细胞过程中，它不会被切割，具有稳定性，不会释放到培养基中，也不会被整合到病毒粒子中。未观察到预测的174 kDa蛋白，而是观察到两种约110 kDa和100 kDa的蛋白。融合蛋白大小的减小是由于有限的糖基化（110 kDa）和未糖基化蛋白（100 kDa）的形成。14k-env蛋白形成寡聚结构，并在病毒感染后暴露于细胞表面。当用表达14K-env融合蛋白的重组病毒接种小鼠时，观察到针对gp160的体液免疫反应。我们的研究结果表明，14k-env蛋白可能具有新的免疫原性特性。
