Plank C, Oberhauser B, Mechtler K, Koch C, Wagner E
Research Institute of Molecular Pathology, Vienna, Austria.
J Biol Chem. 1994 Apr 29;269(17):12918-24.
The process by which viruses destabilize endosomal membranes in an acidification-dependent manner has been mimicked with synthetic peptides that are able to disrupt liposomes, erythrocytes, or endosomes of cultured cells. Peptides containing the 20 amino-terminal amino acid sequence of influenza virus hemagglutinin as well as acidic derivatives showed erythrocyte lysis activity only when peptides were elongated by an amphipathic helix or by carboxyl-terminal dimerization. Interestingly, peptides consisting of the 23 amino-terminal amino acids of influenza virus hemagglutinin were also active in erythrocyte lysis. When peptides were incorporated into DNA complexes that utilize a receptor-mediated endocytosis pathway for uptake into cultured cells, either by ionic interaction with positively charged polylysine-DNA complexes or by a streptavidin-biotin bridge, a strong correlation between pH-specific erythrocyte disruption activity and gene transfer was observed. A high-level expression of luciferase or interleukin-2 was obtained with optimized gene transfer complexes in human melanoma cells and several cell lines.
病毒以酸化依赖方式破坏内体膜的过程已被能够破坏脂质体、红细胞或培养细胞内体的合成肽模拟。含有流感病毒血凝素20个氨基末端氨基酸序列的肽以及酸性衍生物仅在肽通过两亲性螺旋或羧基末端二聚化延长时才显示出红细胞裂解活性。有趣的是，由流感病毒血凝素23个氨基末端氨基酸组成的肽在红细胞裂解中也具有活性。当肽通过与带正电荷的聚赖氨酸-DNA复合物的离子相互作用或通过链霉亲和素-生物素桥掺入利用受体介导的内吞途径摄取到培养细胞中的DNA复合物中时，观察到pH特异性红细胞破坏活性与基因转移之间存在强相关性。在人黑色素瘤细胞和几种细胞系中，使用优化的基因转移复合物获得了荧光素酶或白细胞介素-2的高水平表达。
