Neuropilin-1 and heparan sulfate proteoglycans cooperate in cellular uptake of nanoparticles functionalized by cationic cell-penetrating peptides.
Neuropilin-1 and heparan sulfate proteoglycans cooperate in cellular uptake of nanoparticles functionalized by cationic cell-penetrating peptides.
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DOI：
10.1126/sciadv.1500821
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发表时间：
2015-11
期刊：
影响因子：
13.6
通讯作者：
Ruoslahti E
中科院分区：
文献类型：
--
作者：
Pang HB;Braun GB;Ruoslahti E
两种细胞进入机制在肽介导的细胞内递送中协同作用。细胞穿透肽（CPP）已被广泛用于将纳米材料和其他类型的大分子递送到哺乳动物细胞中用于治疗和诊断用途。结合细胞表面硫酸乙酰肝素（HS）蛋白聚糖的阳离子CPP诱导有效的内吞作用;然而，其他表面受体在此过程中的作用尚不清楚。我们描述了一个HS依赖的途径与C-末端规则（CendR）机制，使肽连接与神经纤毛蛋白-1（NRP 1），已知参与血管生成和血管通透性的细胞表面受体的收敛。NRP 1结合在羧基末端携带阳性残基的肽，这是与阳离子CPP相容的特征，无论是完整的还是蛋白水解加工后的。我们使用CPP和CendR肽，以及HS-和NRP 1结合基序从semaphorins，探讨HS和NRP 1途径的共性和差异。我们发现，CendR-NRP 1相互作用决定了CPP诱导血管通透性的能力。我们还表明，在超微结构水平上，使用一种新的细胞进入同步化方法，HS和NRP 1途径都可以启动一个macropinocytosis-like过程，并可视化这些CPP货物复合物通过各种内体隔室。我们的研究结果为CPP如何利用多种表面受体途径进行细胞内递送提供了新的见解。
Two cell entry mechanisms cooperate in peptide-mediated intracellular delivery. Cell-penetrating peptides (CPPs) have been widely used to deliver nanomaterials and other types of macromolecules into mammalian cells for therapeutic and diagnostic use. Cationic CPPs that bind to heparan sulfate (HS) proteoglycans on the cell surface induce potent endocytosis; however, the role of other surface receptors in this process is unclear. We describe the convergence of an HS-dependent pathway with the C-end rule (CendR) mechanism that enables peptide ligation with neuropilin-1 (NRP1), a cell surface receptor known to be involved in angiogenesis and vascular permeability. NRP1 binds peptides carrying a positive residue at the carboxyl terminus, a feature that is compatible with cationic CPPs, either intact or after proteolytic processing. We used CPP and CendR peptides, as well as HS- and NRP1-binding motifs from semaphorins, to explore the commonalities and differences of the HS and NRP1 pathways. We show that the CendR-NRP1 interaction determines the ability of CPPs to induce vascular permeability. We also show at the ultrastructural level, using a novel cell entry synchronization method, that both the HS and NRP1 pathways can initiate a macropinocytosis-like process and visualize these CPP-cargo complexes going through various endosomal compartments. Our results provide new insights into how CPPs exploit multiple surface receptor pathways for intracellular delivery.