HIV-1 Tat-mediated protein transduction of human brain creatine kinase into PC12 cells.
Epilepsy is characterized by the presence of spontaneous episodes of abnormal neuronal discharges and its pathogenic mechanisms remain poorly understood. Recently, we found that the expression of creatine kinase (CK) was markedly decreased in an epilepsy animal model using proteomic analysis. A human CK gene was fused with a HIV-1 Tat peptide to generate an in-frame Tat-CK fusion protein. The purified Tat-CK 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 Tat-CK fusion protein was stable for 48 h. Moreover, the Tat-CK fusion protein markedly increased endogenous CK activity levels within the cells. These results suggest that Tat-CK provides a strategy for the therapeutic delivery of proteins in various human diseases including the delivery of CK for potential epilepsy treatment.
Jeong
Min Seop
MS
Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University, Chunchon 200-702, Korea.
Kim
Dae Won
DW
Lee
Min Jung
MJ
Lee
Yeom Pyo
YP
Kim
So Young
SY
Lee
Sun Hwa
SH
Jang
Sang Ho
SH
Lee
Kil Soo
KS
Park
Jinseu
J
Kang
Tae-Cheon
TC
Cho
Sung-Woo
SW
Kwon
Oh-Shin
OS
Eum
Won Sik
WS
Choi
Soo Young
SY
D000818:Animals; D003001:Cloning, Molecular; D052276:Creatine Kinase, BB Form / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D016716:PC12 Cells; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000235:genetics / Q000302:isolation & purification; D014161:Transduction, Genetic / Q000379:methods*; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics* / Q000302:isolation & purification
D011993:Recombinant Fusion Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus; D052276:Creatine Kinase, BB Form