Differences in the post-translational modifications of human papillomavirus type 6b major capsid protein expressed from a baculovirus system compared with a vaccinia virus system
Differences in the post-translational modifications of human papillomavirus type 6b major capsid protein expressed from a baculovirus system compared with a vaccinia virus system
复制标题
DOI：
10.1042/ba20000001
复制
发表时间：
2000-08-01
影响因子：
2.8
通讯作者：
Fernando, GJP
中科院分区：
文献类型：
--
作者：
Fang, NX;Frazer, IH;Fernando, GJP
目前正在研究病毒样颗粒（VLP）在人类病毒感染的疫苗中的作用。有不同的重组蛋白表达系统可用于获得疫苗接种所需的VLP制剂。然而，所获得的重组蛋白的翻译后修饰的差异及其在疫苗中引发抗病毒应答的功效的差异尚未充分确定。在这项研究中，我们比较了人乳头瘤病毒6 b型主要衣壳蛋白L1的翻译后修饰，使用重组杆状病毒（rBV）在Sf 9中表达的HPV 6 bL 1（草地贪夜蛾）昆虫细胞，其中使用重组牛痘病毒（rVV）在CV-1肾上皮细胞中表达蛋白质，生物合成标记的rBV表达的HPV 6 bL 1的双向凝胶电泳显示了该蛋白的几种后修饰变体，而rVV表达的HPV 6 bL 1仅显示少数变异。磷酸化检测到苏氨酸和丝氨酸残基的L1表达的rBV相比，丝氨酸残基的磷酸化仅为L1表达的rVV。用掺入[H-3]甘露糖和[H-3]半乳糖的rBV表达HPV 6 bL 1，而用仅掺入[H-3]半乳糖的rVV表达HPV 6 bL 1。我们得出结论，重组HPV 6 bL 1的翻译后修饰可以根据其表达所用的系统而有所不同。由于重组L1蛋白是一种潜在的人类疫苗候选者，因此需要研究所观察到的翻译后修饰对L1 VLP免疫原性的影响。
Virus-like particles (VLPs) are being currently investigated in vaccines against viral infections in humans. There are different recombinant-protein-expression systems available for obtaining the necessary VLP preparation for vaccination. However, the differences in post-translational modifications of the recombinant proteins obtained and their differences in efficacy in eliciting an anti-viral response in vaccines are not well established. In this study we have compared the posttranslational modifications of human papillomavirus type-6b major capsid protein L1 (HPV 6bL1) expressed using recombinant baculovirus (rBV) in Sf9 (Spodoptera frugiperda) insect cells, with the protein expressed using recombinant vaccinia virus (rVV) in CV-1 kidney epithelial cells, Two-dimensional gel electrophoresis of biosynthetically labelled rBV-expressed HPV 6bL1 showed several post-translationally modified variants of the protein, whereas rVV-expressed HPV 6bL1 showed only a few variants. Phosphorylations were detected at threonine and serine residues for the L1 expressed from rBV compared with phosphorylation at serine residues only for the L1 expressed from rVV. HPV 6bL1 expressed using rBV incorporated [H-3]mannose and [H-3]galactose, whereas HPV 6bL1 expressed using rVV incorporated only [H-3]galactose. We conclude that post-translational modification of recombinant HPV 6bL1 can differ according to the system used for its expression. Since recombinant L1 protein is a potential human-vaccine candidate, the implication of the observed differences in post-translational modifications on immunogenicity of L1 VLPs warrants investigation.