PMID: 11918970Mar 29, 2002Paper

TAT-mediated delivery of human glutamate dehydrogenase into PC12 cells

Neurochemistry International
Hye-Young YoonSoo Young Choi

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.

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Citations

Jul 13, 2006·Annual Review of Biomedical Engineering·Vladimir P Torchilin
Oct 16, 2004·Molecular and Cellular Neurosciences·Gunnar P H Dietz, Mathias Bähr
Apr 16, 2014·Journal of Drug Targeting·Tzyy-Harn YehLi-Jiuan Shen
Dec 6, 2005·Brain Research Bulletin·Gunnar P H Dietz, Mathias Bähr
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Feb 22, 2005·Advanced Drug Delivery Reviews·Bhawna GuptaVladimir P Torchilin
Jul 5, 2003·Molecular Therapy : the Journal of the American Society of Gene Therapy·Siobhan M CashmanRajendra Kumar-Singh
Mar 30, 2010·Pharmaceuticals·Sara TrabuloMaria C Pedroso De Lima
Mar 17, 2006·IUBMB Life·Alexandre KerkisIrina Kerkis
Jul 3, 2004·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Alexandre KerkisTetsuo Yamane
Oct 1, 2008·Drug Discovery Today. Technologies·Vladimir Torchilin

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