Cytosolic delivery of macromolecules 4. Head group-dependent membrane permeabilization by pH-sensitive detergents.
Cytosolic delivery of macromolecules 4. Head group-dependent membrane permeabilization by pH-sensitive detergents.
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大分子的胞质传递 4. pH 敏感洗涤剂的头基依赖性膜透化。
DOI：
10.1016/j.jconrel.2005.04.010
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Cho,MooJ
中科院分区：
文献类型：
--
作者：
Asokan,Aravind;Cho,MooJ
三个叔胺基洗涤剂具有零个，一个，或两个羟基在其头部基团的不同位置，其特征在于它们的能力，以促进大分子的胞质传递。脂质结构的临界胶束浓度（CMC）和膜结合pKa值随着头基极性的增加而增加，范围分别为1-5 μM和5.9 - 6.3。荧光共振能量转移（FRET）和钙黄绿素泄漏实验表明，当胺基质子化引入-OH部分的去污剂头基团增强其相互作用和透化阴离子，内体模拟囊泡的能力。进一步评价了基于二乙醇胺的脂质（DEL）的不同制剂的pH依赖性溶血活性和促进体外大分子的胞质递送的能力。完整的脂质体含有DEL在其最大限度的掺入效率低于DEL含胶束在促进血红蛋白泄漏从人红细胞在酸性pH值。在HeLa细胞中，DEL含洗涤剂胶束促进有效的胞质释放的内吞的大分子，如荧光素标记的葡聚糖的MW 10 kDa。基于增强型绿色荧光蛋白（EGFP）的反义介导的上调的功能测定进一步证实了这一观察结果。总之，我们的研究结果强调了极性头基和pH敏感性洗涤剂的胶束结构在介导内体透化和大分子的有效胞质递送中的关键作用。
Three tertiary amine-based detergents with zero, one, or two hydroxyl groups at various positions in their head group were characterized for their ability to promote the cytosolic delivery of macromolecules. Critical micellar concentrations (CMC) and membrane-bound pKa values of the lipid constructs increased with increasing head group polarity, ranging from 1–5 μM and 5.9 to 6.3, respectively. Fluorescence resonance energy transfer (FRET) and calcein leakage experiments revealed that when the amine group is protonated introduction of –OH moieties to detergent head groups enhanced their ability to interact with and permeabilize anionic, endosome-mimicking vesicles. Different formulations of a diethanolamine-based lipid (DEL) were further evaluated for pH-dependent hemolytic activity and ability to promote cytosolic delivery of macromolecules in vitro. Intact liposomes containing DEL at its maximum limit of incorporation were less efficient than DEL-containing micelles in promoting hemoglobin leakage from human erythrocytes at acidic pH. In HeLa cells, DEL-containing detergent micelles facilitated efficient cytosolic release of endocytosed macromolecules such as fluorescein-labeled dextran of MW 10 kDa. This observation was further corroborated by a functional assay based on antisense-mediated up-regulation of enhanced green fluorescent protein (EGFP). Taken together, our findings emphasize the key role of polar head groups and micellar architecture of pH-sensitive detergents in mediating endosomal permeabilization and the efficient cytosolic delivery of macromolecules.