Binding of cell-penetrating penetratin peptides to plasma membrane vesicles correlates directly with cellular uptake
Binding of cell-penetrating penetratin peptides to plasma membrane vesicles correlates directly with cellular uptake
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DOI：
10.1016/j.bbamem.2011.03.011
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发表时间：
2011-07-01
影响因子：
3.4
通讯作者：
Esbjoerner, Elin K.
中科院分区：
文献类型：
--
作者：
Amand, Helene L.;Bostrom, Carolina L.;Esbjoerner, Elin K.
细胞穿透肽（CPP）主要通过内吞作用进入细胞内，并具有将大分子货物递送到细胞内的能力。尽管已经描述了各种内吞途径的参与，但仍不清楚哪些相互作用参与引发摄取反应，以及对特定细胞表面组分的亲和力在多大程度上可以决定特定CPP的效率。以前的生物物理学研究CPP和脂囊泡或细胞表面蛋白聚糖的可溶性糖模拟物，两个最常见的建议CPP结合靶点之间的相互作用，还没有允许建立定量相关性。在这里，我们探索使用的质膜囊泡（PMVs）来自培养的哺乳动物细胞作为细胞表面模型在生物物理实验。此外，我们使用CPP穿透素和分别富含精氨酸和赖氨酸的两种类似物检查了对PMV的亲和力和摄取到活细胞中之间的关系。我们表明，使用离心沉淀PMVs，在颗粒级分中的肽的量与细胞内化的程度线性相关，并且穿透素的所有精氨酸和所有赖氨酸变体的相对效率可以归因于它们各自的细胞表面亲和力。我们的数据显示了富含精氨酸和赖氨酸的穿透素变体之间的差异，这些差异之前在使用脂囊泡的研究中尚未得到解释。我们的数据还表明，更大的差异，结合亲和力PMVs比肝素，一种常用的细胞表面蛋白聚糖模拟。总而言之，这表明CPP的细胞表面相互作用取决于几个细胞表面部分及其在质膜上的分子组织。(C)2011 Elsevier BM. All rights reserved.
Cell-penetrating peptides (CPPs) gain access to intracellular compartments mainly via endocytosis and have capacity to deliver macromolecular cargo into cells. Although the involvement of various endocytic routes has been described it is still unclear which interactions are involved in eliciting an uptake response and to what extent affinity for particular cell surface components may determine the efficiency of a particular CPP. Previous biophysical studies of the interaction between CPPs and either lipid vesicles or soluble sugar-mimics of cell surface proteoglycans, the two most commonly suggested CPP binding targets, have not allowed quantitative correlations to be established. We here explore the use of plasma membrane vesicles (PMVs) derived from cultured mammalian cells as cell surface models in biophysical experiments. Further, we examine the relationship between affinity for PMVs and uptake into live cells using the CPP penetratin and two analogs enriched in arginines and lysines respectively. We show, using centrifugation to sediment PMVs, that the amount of peptide in the pellet fraction correlates linearly with the degree of cell internalization and that the relative efficiency of all-arginine and all-lysine variants of penetratin can be ascribed to their respective cell surface affinities. Our data show differences between arginine- and lysine-rich variants of penetratin that has not been previously accounted for in studies using lipid vesicles. Our data also indicate greater differences in binding affinity to PMVs than to heparin, a commonly used cell surface proteoglycan mimic. Taken together, this suggests that the cell surface interactions of CPPs are dependent on several cell surface moieties and their molecular organization on the plasma membrane. (C) 2011 Elsevier BM. All rights reserved.