In vivo protein transduction: Biologically active intact PEP-1-superoxide dismutase fusion protein efficiently protects against ischemic insult
In vivo protein transduction: Biologically active intact PEP-1-superoxide dismutase fusion protein efficiently protects against ischemic insult
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DOI：
10.1016/j.freeradbiomed.2004.07.028
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发表时间：
2004-11-15
影响因子：
7.4
通讯作者：
Choi, SY
中科院分区：
文献类型：
--
作者：
Eum, WS;Kim, DW;Choi, SY
活性氧与短暂性局灶性脑缺血再灌注损伤有关。抗氧化酶铜锌超氧化物歧化酶是细胞对抗缺血后ROS损伤的主要途径之一。最近，我们报道了变性TAT-SOD融合蛋白被转导到细胞和皮肤组织中。此外，PEP-1多肽具有21个氨基酸残基，是一种已知的载体多肽，可在体外和体内运送全长天然蛋白。本研究旨在探讨PEP-1-SOD融合蛋白对脑缺血损伤的保护作用。将人超氧化物歧化酶基因与PEP-1多肽融合到细菌表达载体中，获得了PEP-1-SOD基因框内融合蛋白。表达和纯化的融合蛋白在体外和体内均能有效转导，具有天然的蛋白结构。免疫组织化学分析显示，PEP-1-SOD经腹腔注射(i.p)。进入老鼠体内可以进入大脑神经元。当将PEP-1-SOD融合蛋白注射到沙土鼠体内时，可以防止由短暂性前脑缺血引起的海马神经细胞死亡。这些结果表明，具有生物活性的PEP-1-SOD的完整形式为治疗与这种抗氧化酶或ROS相关的各种人类疾病提供了一种更有效的策略，包括中风。(C)2004 Elsevier Inc.保留所有权利。
Reactive oxygen species (ROS) are implicated in reperfusion injury after transient focal cerebral ischemia. The antioxidant enzyme Cu,Zn-superoxide dismutase (SOD) is one of the major means by which cells counteract the deleterious effects of ROS after ischemia. Recently, we reported that denatured Tat-SOD fusion protein is transduced into cells and skin tissue. Moreover, PEP-1 peptide, which has 21 amino acid residues, is a known carrier peptide that delivers full-length native proteins in vitro and in vivo. In the present study, we investigated the protective effects of PEP-1-SOD fusion protein after ischemic insult. A human SOD gene was fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-SOD fusion protein. The expressed and purified fusion proteins were efficiently transduced both in vitro and in vivo with a native protein structure. Immunohistochemical analysis revealed that PEP-1-SOD injected intraperitoneally (i.p.) into mice can have access into brain neurons. When i.p.-injected into gerbils, PEP-1-SOD fusion proteins prevented neuronal cell death in the hippocampus caused by transient forebrain ischemia. These results suggest that the biologically active intact forms of PEP-1-SOD provide a more efficient strategy for therapeutic delivery in various human diseases related to this antioxidant enzyme or to ROS, including stroke. (C) 2004 Elsevier Inc. All rights reserved.