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首页> 外文期刊>Biochemistry >Self-promoted cellular uptake of peptide/DNA transfection complexes
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Self-promoted cellular uptake of peptide/DNA transfection complexes

机译：自我促进的肽/ DNA转染复合物的摄取

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摘要

The designed alpha-helical amphipathic peptide LAH4 assembles several properties, which makes it an interesting candidate as a gene-delivery vehicle. Besides being short and soluble in aqueous solutions, LAH4 presents cationic residues, which allow for efficient complexation of DNA. In addition, this peptide is poorly hemolytic at neutral pH, while it is able to destabilize biological membranes in acidic conditions. In this study, the structure of the peptide/DNA transfection complex was examined by circular dichroism and solid-state nuclear magnetic resonance spectroscopies and the thermodynamics of its formation and disassembly was monitored in a quantitative manner as a function of pH by isothermal titration calorimetry. Notably, the number of peptides within the complex considerably decreases upon acidification of the medium. This observation has direct and important consequences for the mechanism of action because the acidification of the endosome results in high local concentrations of free peptide in this organelle. Thus, these peptides become available to interact with the endosomal membranes and thereby responsible for the delivery of the transfection complex to the cytoplasm. When these data are taken together, they indicate a dual role of the peptide during the transfection process, namely, DNA complexation and membrane permeabilization.
机译：设计的α-螺旋肌肤肽Lah4组装了几种性质，这使其成为一个有趣的候选者作为基因输送载体。除了短期和溶于水溶液之外，LAH4呈阳离子残留物，允许有效络合DNA。此外，该肽在中性pH下溶血性差，而它能够使生物膜变得破坏酸性条件。在该研究中，通过圆形二色性和固态核磁共振谱检查肽/ DNA转染复合物的结构，并以定量方式通过等温滴定热量测量其形成的热力学和拆卸的热力学。值得注意的是，在培养基酸化时，复合物内的肽的数量显着降低。这种观察结果对动作机制具有直接和重要的后果，因为内部体的酸化导致该细胞器中的高局部肽的游离肽。因此，这些肽可用于与内体膜相互作用，从而负责递送转染复合物到细胞质。当这些数据占据在一起时，它们表示肽在转染过程中的双重作用，即DNA络合和膜渗透。

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