On the mechanism of non-endosomial peptide-mediated cellular delivery of nucleic acids
On the mechanism of non-endosomial peptide-mediated cellular delivery of nucleic acids
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DOI：
10.1016/j.bbamem.2004.09.010
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发表时间：
2004-12-15
影响因子：
3.4
通讯作者：
Heitz, F
中科院分区：
文献类型：
--
作者：
Deshayes, S;Gerbal-Chaloin, S;Heitz, F
最近，我们描述了一种新的策略，用于将核酸递送到哺乳动物细胞中，基于称为MPG的27个残基的两亲性肽，其是基于衍生自与核定位序列相关的融合序列的疏水结构域设计的，并通过接头分离。这种肽载体构成了在培养细胞中递送核酸的有力工具，而不需要任何共价偶联。我们已经研究了MPG的构象状态，在其自由形式和复杂的货物，以及其与磷脂相互作用的能力，并调查了这些相互作用的结构后果。尽管它的相似性，类似设计的细胞穿透肽Pep-1，MPG的行为显着不同的构象的观点。圆二色性（CD）分析揭示了从非结构化的β-折叠构象与磷脂相互作用后的过渡。我们建议，跨膜过程涉及到一个短暂的跨膜孔样结构的形成。MPG的部分构象变化与其货物的复合物的形成有关，并且在与细胞膜缔合后发生片层含量的增加。(C)2004 Elsevier B. V.保留所有权利。
Recently, we described a new strategy for the delivery of nucleic acids into mammalian cells, based on an amphipathic peptide of 27 residues called MPG, which was designed on the basis of a hydrophobic domain derived from a fusion sequence associated with a nuclear localization sequence and separated by a linker. This peptide carrier constitutes a powerful tool for the delivery of nucleic acids in cultured cells, without requiring any covalent coupling. We have examined the conformational states of MPG in its free form and complexed with a cargo, as well as its ability to interact with phospholipids, and have investigated the structural consequences of these interactions. In spite of its similarity to the similarly designed cell-penetrating peptide Pep-1, MPG behaves significantly differently from the conformational point of view. Circular dichroism (CD) analysis reveals a transition from a nonstructured to a beta-sheet conformation upon interaction with phospholipids. We propose that the membrane crossing process involves formation of a transient transmembrane pore-like structure. Partial conformational change of MPG is associated with formation of a complex with its cargo, and an increase in sheet content occurs upon association with the cell membrane. (C) 2004 Elsevier B.V. All rights reserved.