title: TAT-mediated delivery of human glutamate dehydrogenase into PC12 cells.
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.
authors.affiliation: Department of Biochemistry, University of Ulsan College of Medicine, Seoul, South Korea.
authors.forename: Hye-Young
authors.initials: HY
authors.lastname: Yoon
authors.forename: Sung Ho
authors.initials: SH
authors.lastname: Lee
authors.forename: Sung-Woo
authors.initials: SW
authors.lastname: Cho
authors.forename: Jong Eun
authors.initials: JE
authors.lastname: Lee
authors.forename: Chang Sik
authors.initials: CS
authors.lastname: Yoon
authors.forename: Jinseu
authors.initials: J
authors.lastname: Park
authors.forename: Tae Ue
authors.initials: TU
authors.lastname: Kim
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Choi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015696:Gene Products, tat / Q000008:administration & dosage*; D005969:Glutamate Dehydrogenase / Q000008:administration & dosage / Q000378:metabolism*; D006678:HIV / Q000378:metabolism*; D006801:Humans; D016716:PC12 Cells; D051381:Rats; D054322:tat Gene Products, Human Immunodeficiency Virus
chemical_list: D015696:Gene Products, tat; D054322:tat Gene Products, Human Immunodeficiency Virus; D005969:Glutamate Dehydrogenase