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  • Title: Tat-mediated protein transduction of human brain pyridoxal kinase into PC12 cells.
    Author: Kim DW, Kim CK, Choi SH, Choi HS, Kim SY, An JJ, Lee SR, Lee SH, Kwon OS, Kang TC, Won MH, Cho YJ, Cho SW, Kang JH, Kim TY, Lee KS, Park J, Eum WS, Choi SY.
    Journal: Biochimie; 2005 May; 87(5):481-7. PubMed ID: 15820755.
    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.
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