Peptide Derived from HIV-1 TAT Protein Destabilizes a Monolayer of Endothelial Cells in an in Vitro Model of the Blood-Brain Barrier and Allows Permeation of High Molecular Weight Proteins
Peptide Derived from HIV-1 TAT Protein Destabilizes a Monolayer of Endothelial Cells in an in Vitro Model of the Blood-Brain Barrier and Allows Permeation of High Molecular Weight Proteins
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DOI：
10.1074/jbc.m112.395384
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发表时间：
2012-12-28
影响因子：
4.8
通讯作者：
Shechter, Yoram
中科院分区：
文献类型：
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作者：
Cooper, Itzik;Sasson, Keren;Shechter, Yoram
大多数化学治疗剂是血脑屏障（BBB）不渗透剂。HIV-1衍生的达特蛋白变体含有跨膜结构域，这可能使它们能够穿过BBB并到达大脑。在这里，我们合成了CAYGRK-KRRQRRR，一种含有连接到跨膜结构域（C-TAT肽）的N末端的半胱氨酸部分的肽，并研究了其在体外BBB模型中的作用，我们发现该模型反映了受体非依赖性途径的渗透。将脑毛细血管内皮细胞单层与0.3-0.6 μ mol/ml的这种C-TAT肽孵育1-2小时，使脑毛细血管内皮细胞单层不稳定，并引入包括高分子量蛋白质的不渗透治疗剂基本上穿透它的能力。C-TAT肽1位的半胱氨酰部分在很大程度上有助于不渗透物质的去稳定效力和渗透效力。使用肝素可逆转不稳定效应。总之，在体外BBB模型中发现了允许不渗透的低分子量和高分子量物质从腔侧（血液）进入近腔侧（脑）的显著增加的实验条件，该模型反映了通过受体非依赖性途径的体内蛋白质渗透性。
Most chemotherapeutic agents are blood-brain barrier (BBB) impermeants. HIV-1-derived TAT protein variants contain a transmembrane domain, which may enable them to cross the BBB and reach the brain. Here we synthesized CAYGRK-KRRQRRR, a peptide containing a cysteine moiety attached to the N terminus of the transmembrane domain (C-TAT peptide), and studied its effects in an in vitro BBB model, which we found to reflect penetration by a receptor-independent pathway. Incubation of the brain capillary endothelial cell monolayer with 0.3-0.6 mu mol/ml of this C-TAT peptide, for a period of 1-2 h, destabilizes brain capillary endothelial cell monolayer and introduces the ability of impermeant therapeutic agents including high molecular weight proteins to penetrate it substantially. The cysteinyl moiety at position 1 of the C-TAT peptide contributes largely to the destabilizing potency and the penetration efficacy of impermeant substances. The destabilizing effect was reversed using heparin. In summary, experimental conditions allowing a significant increase in entry of impermeant low and high molecular weight substances from the luminal (blood) to the abluminal side (brain) were found in an in vitro BBB model reflecting in vivo protein penetrability by a receptor-independent pathway.