Human brain pyridoxal-5'-phosphate phosphatase (PLPP):protein transduction of PEP-1-PLPP into PC12 cells.
Pyridoxal-5'-phosphate phosphatase (PLPP) catalyzes the dephosphorylation of pyridoxal-5'-phosphate (PLP). A human brain PLPP gene was fused with a PEP-1 peptide and produced a genetic in-frame PEP-1-PLPP fusion protein. The purified PEP-1-PLPP fusion protein was efficiently transduced into PC12 cells in a time- and dose-dependent manner when added exogenously to culture media. Once inside the cells, the transduced PEP-1-PLPP fusion protein was stable for 36 h. The concentration of PLP was markedly decreased by the addition of exogenous PEP-1-PLPP to media pretreated with the vitamin B(6) precursors; pyridoxine, pyridoxal kinase and pyridoxine-5'-phosphate oxidase into cells. The results suggest that the transduction of the PEP-1-PLPP fusion protein can be one mode of PLP level regulation, and to replenish this enzyme in the various neurological disorders related to vitamin B(6).
Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University, Chuncheon, Korea.
Yeom Pyo
YP
Lee
Dae Won
DW
Kim
Min Jung
MJ
Lee
Min Seop
MS
Jeong
So Young
SY
Kim
Sun Hwa
SH
Lee
Sang Ho
SH
Jang
Jinseu
J
Park
Tae-Cheon
TC
Kang
Moo Ho
MH
Won
Sung-Woo
SW
Cho
Oh-Shin
OS
Kwon
Won Sik
WS
Eum
Soo Young
SY
Choi
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology*; D003543:Cysteamine / Q000031:analogs & derivatives* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D016716:PC12 Cells; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
C503924:Pep-1 peptide; D010455:Peptides; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D003543:Cysteamine; C027138:pyridoxine phosphate phosphatase; D010744:Phosphoric Monoester Hydrolases