Abstract：
: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.
journal_name
Neurochem Intjournal_title
Neurochemistry internationalauthors
Yoon HY,Lee SH,Cho SW,Lee JE,Yoon CS,Park J,Kim TU,Choi SYdoi
10.1016/s0197-0186(01)00138-3subject
Has Abstractpub_date
2002-07-01 00:00:00pages
37-42issue
1eissn
0197-0186issn
1872-9754pii
S0197018601001383journal_volume
41pub_type
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