title: Tat-mediated protein transduction of human brain pyridoxal kinase into PC12 cells.
abstract: Pyridoxal kinase (PK) catalyses the phosphorylation of vitamin B6 to pyridoxal-5'-phosphate (PLP). A human brain PK gene was fused with a gene fragment encoding the HIV-1 Tat protein transduction domain (RKKRRQRRR) in a bacterial expression vector to produce a genetic in-frame Tat-PK fusion protein. The expressed and purified Tat-PK fusion proteins transduced efficiently into PC12 cells in a time- and dose-dependent manner when added exogenously in culture media. Once inside the cells, the transduced Tat-PK proteins showed catalytic activity and are stable for 48 h. The intracellular concentration of PLP, which is known as a biologically active form of vitamin B6, was increased by pre-treatment of Tat-PK to the PC12 cells. Those results suggest that the transduction of Tat-PK fusion protein can be one of the ways to regulate the PLP level and to replenish this enzyme in the various neurological disorders related to vitamin B6.
authors.affiliation: Department of Genetic Engineering and Research Institute for Bioscience and Biotechnology, Hallym University, Chunchon 200-702, South Korea.
authors.forename: Dae Won
authors.initials: DW
authors.lastname: Kim
authors.forename: Chung Kwon
authors.initials: CK
authors.lastname: Kim
authors.forename: Soo Hyun
authors.initials: SH
authors.lastname: Choi
authors.forename: Hee Soon
authors.initials: HS
authors.lastname: Choi
authors.forename: So Young
authors.initials: SY
authors.lastname: Kim
authors.forename: Jae Jin
authors.initials: JJ
authors.lastname: An
authors.forename: Seung Ree
authors.initials: SR
authors.lastname: Lee
authors.forename: Sun Hwa
authors.initials: SH
authors.lastname: Lee
authors.forename: Oh-Shin
authors.initials: OS
authors.lastname: Kwon
authors.forename: Tae-Cheon
authors.initials: TC
authors.lastname: Kang
authors.forename: Moo Ho
authors.initials: MH
authors.lastname: Won
authors.forename: Yong Joon
authors.initials: YJ
authors.lastname: Cho
authors.forename: Sung-Woo
authors.initials: SW
authors.lastname: Cho
authors.forename: Jung Hoon
authors.initials: JH
authors.lastname: Kang
authors.forename: Tae Yoon
authors.initials: TY
authors.lastname: Kim
authors.forename: Kil Soo
authors.initials: KS
authors.lastname: Lee
authors.forename: Jinseu
authors.initials: J
authors.lastname: Park
authors.forename: Won Sik
authors.initials: WS
authors.lastname: Eum
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Choi
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology*; D015696:Gene Products, tat / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D016716:PC12 Cells / Q000378:metabolism; D011731:Pyridoxal Kinase / Q000235:genetics*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: D015696:Gene Products, tat; D011993:Recombinant Fusion Proteins; D011731:Pyridoxal Kinase