Full-length proteins attached to the HIV tat protein transduction domain are neither transduced between cells, nor exhibit enhanced immunogenicity
Full-length proteins attached to the HIV tat protein transduction domain are neither transduced between cells, nor exhibit enhanced immunogenicity
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DOI：
10.1038/sj.gt.3301819
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发表时间：
2002-11-01
期刊：
影响因子：
5.1
通讯作者：
Whitton, JL
中科院分区：
文献类型：
--
作者：
Leifert, JA;Harkins, S;Whitton, JL
一些蛋白质已经被赋予了不寻常的能力，以受体非依赖性和温度非依赖性的方式跨细胞膜易位，这种活性已被映射到一个高度碱性的残基系列，目前被称为“蛋白质转导结构域”（PTD）。这种易位属性，如果真实的话，将是有价值的基因治疗和疫苗接种的目的。我们已经评估了来自人类免疫缺陷病毒1型（HIV）达特蛋白的PTD，并且我们得出结论，当从头合成时，（1）HIV达特PTD不会增强其所连接的全长蛋白的免疫原性;（2）通过仔细的定量分析判断，HIV达特PTD不会引起所连接的标记蛋白的细胞间转移。从我们的数据和对已发表材料的回顾中，我们认为，与目前的教条相反，几乎没有证据表明这些所谓的易位蛋白质可以在活细胞之间移动。此外，我们认为PTDs不起增强易位的作用，而仅仅是增加与细胞表面的结合;在这种情况下，术语“蛋白转导结构域”和相关的首字母缩略词是应该放弃的用词不当。我们的结论解释了为什么最引人注目的示范PTD疗效已获得使用固定的细胞和/或变性蛋白质，并有明显的基因治疗和疫苗接种的影响。
Several proteins have been accorded the unusual ability to translocate across cell membranes in a receptor-independent and temperature-independent manner, and this activity has been mapped to a highly basic series of residues currently termed a 'protein transduction domain' (PTD). This translocatory attribute, if authentic, would be valuable for purposes of gene therapy and vaccination. We have evaluated the PTD from the human immunodeficiency virus type 1 (HIV) tat protein and we conclude that, when synthesized de novo, (1) the HIV tat PTD does not enhance the immunogenicity of a full-length protein to which it is tethered; and (2) the HIV tat PTD does not cause intercellular transfer of an attached marker protein, as judged by careful quantitative analyses. From our data, and from a review of published materials, we suggest that contrary to current dogma there is little evidence that these supposedly translocatory proteins can move between live cells. Furthermore, we suggest that PTDs do not act to enhance translocation, but instead merely to increase binding to the cell surface; in which case, the term 'protein transduction domain, and the related acronym, are misnomers which should be abandoned. Our conclusions explain why the most dramatic demonstrations of PTD efficacy have been obtained using fixed cells and/or denatured proteins, and have obvious implications for gene therapy and vaccination.