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首页> 外文期刊>Neurochemistry International: The International Journal for the Rapid Publication of Critical Reviews, Preliminary and Original Research Communications in Neurochemistry >TAT-mediated delivery of human glutamate dehydrogenase into PC12 cells.
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TAT-mediated delivery of human glutamate dehydrogenase into PC12 cells.

机译：TAT介导的人谷氨酸脱氢酶向PC12细胞的传递。

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摘要

Human glutamate dehydrogenase (GDH) gene was fused with a gene fragment encoding the nine amino acid (RKKRRQRRR) protein transduction domain of human immunodeficiency virus TAT protein in bacterial expression vector to produce genetic in-frame TAT-GDH fusion protein. The TAT-GDH protein can enter PC12 cells efficiently when added exogenously in culture media as determined by Western blot analysis and enzyme activities. Once inside the cells, the transduced denatured TAT-GDH protein showed a full activity of GDH indicating that the TAT-GDH fusion protein was correctly refolded after delivery into cells and the activities of GDH in the TAT-GDH fusion protein was not affected by the addition of the TAT sequence. TAT-GDH fusion protein and TAT itself showed no cytotoxicity in PC12 cells. Although the exact mechanism of transduction across a membrane remains unclear, the transduction activity of TAT-GDH into PC12 cells may suggest new possibilities for direct delivery of GDH into the patients with the GDH-deficient disorders.
机译：将人谷氨酸脱氢酶（GDH）基因与编码人免疫缺陷病毒TAT蛋白的9个氨基酸（RKKRRQRRR）蛋白转导域的基因片段在细菌表达载体中融合，以产生遗传框内TAT-GDH融合蛋白。如通过蛋白质印迹分析和酶活性确定，当外源添加到培养基中时，TAT-GDH蛋白可以有效进入PC12细胞。一旦进入细胞，转导的变性TAT-GDH蛋白就显示出GDH的完整活性，这表明TAT-GDH融合蛋白在递送入细胞后已正确地重折叠，并且TAT-GDH融合蛋白中的GDH活性不受此影响。 TAT序列的添加。 TAT-GDH融合蛋白和TAT本身对PC12细胞无细胞毒性。尽管跨膜的确切转导机制尚不清楚，但TAT-GDH向PC12细胞的转导活性可能为将GDH直接递送至GDH缺乏症患者提供了新的可能性。

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