Complexes of DNA with cationic peptides: Conditions of formation and factors effecting internalization by mammalian cells
Complexes of DNA with cationic peptides: Conditions of formation and factors effecting internalization by mammalian cells
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DOI：
10.1134/s0006297906120108
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发表时间：
2006-12-01
影响因子：
2.8
通讯作者：
Orlov, S. V.
中科院分区：
文献类型：
--
作者：
Dizhe, E. B.;Ignatovich, I. A.;Orlov, S. V.
本文研究了DNA/K_8复合物的形成条件，并与人免疫缺陷病毒（1型）转录因子达特（达特肽）的富含精氨酸片段（47-57）的DNA复合物进行了比较，分析了影响DNA/K_8复合物进入哺乳动物细胞的因素。首次研究了带正电荷的DNA/K8复合物的化学计量。DNA-K8相互作用的非合作性质。已经表明，沿着这种复合物的正电荷，培养基中存在过量的游离K8肽是用DNA/K8复合物进行细胞转染的最大效率的必要条件。游离K8肽对DNA/K8复合物转染哺乳动物细胞效率的刺激作用可能是通过阳离子肽K8与细胞表面带负电荷的蛋白聚糖的相互作用介导的，这导致保护DNA/K8复合物免受细胞硫酸乙酰肝素的破坏。然而，游离阳离子肽的保护作用不仅取决于它们的正电荷，还取决于肽的一级结构。与具有基于聚-L-赖氨酸的分子缀合物的DNA复合物所获得的结果相反，DNA/K8复合物的聚集导致转移基因的表达显著增加。
This work was devoted to the study of conditions of the formation of DNA/K8 complex and analysis of factors effecting the entry of DNA/K8 complex into mammalian cells in comparison with DNA complexes with arginine-rich fragment (47-57) of human immunodeficiency virus (type 1) transcription factor Tat (Tat peptide). The stoichiometry of positively charged DNA/K8 complexes has been studied for the first time. Non-cooperative character of DNA-K8 interaction was revealed. It has been shown that along with the positive charge of such complexes, the presence of an excess of free K8 peptide in the culture medium is a necessary condition for maximal efficiency of cell transfection with DNA/K8 complexes. A stimulatory effect of free K8 peptide on the efficiency of mammalian cell transfection by DNA/K8 complexes is likely to be mediated by the interactions of cationic peptide K8 with negatively charged proteoglycans on the cell surface, which leads to protection of DNA/K8 complexes from disruption by cellular heparan sulfates. However, the protective role of free cationic peptides depends not only on their positive charge, but also on the primary structure of the peptide. In contrast with the results obtained for DNA complexes with molecular conjugates based on poly-L-lysine, the aggregation of DNA/K8 complexes leads to a significant increase in the expression of transferred gene.