Tat-mediated protein transduction of human brain pyridoxal kinase into PC12 cells

Dae Won Kim; Chung Kwon Kim; Soo Hyun Choi; Hee Soon Choi; So Young Kim; Jae Jin An; Seung Ree Lee; Sun Hwa Lee; Oh Shin Kwon; Tae Cheon Kang; Moo Ho Won; Yong Joon Cho; Sung Woo Cho; Jung Hoon Kang; Tae Yoon Kim; Kil Soo Lee; Jinseu Park; Won Sik Eum; Soo Young Choi

doi:10.1016/j.biochi.2004.12.004

Tat-mediated protein transduction of human brain pyridoxal kinase into PC12 cells

Dae Won Kim, Chung Kwon Kim, Soo Hyun Choi, Hee Soon Choi, So Young Kim, Jae Jin An, Seung Ree Lee, Sun Hwa Lee, Oh Shin Kwon, Tae Cheon Kang, Moo Ho Won, Yong Joon Cho, Sung Woo Cho, Jung Hoon Kang, Tae Yoon Kim, Kil Soo Lee, Jinseu Park, Won Sik Eum, Soo Young Choi

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

Abstract

Pyridoxal kinase (PK) catalyses the phosphorylation of vitamin B ₆ 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.

Original language	English
Pages (from-to)	481-487
Number of pages	7
Journal	Biochimie
Volume	87
Issue number	5
DOIs	https://doi.org/10.1016/j.biochi.2004.12.004
State	Published - May 2005

Keywords

Brain pyridoxal kinase
HIV-1 Tat
Protein therapy
Pyridoxal-5′-phosphate
Transduction

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.biochi.2004.12.004

Cite this

Kim, D. W., Kim, C. K., Choi, S. H., Choi, H. S., Kim, S. Y., An, J. J., Lee, S. R., Lee, S. H., Kwon, O. S., Kang, T. C., Won, M. H., Cho, Y. J., Cho, S. W., Kang, J. H., Kim, T. Y., Lee, K. S., Park, J., Eum, W. S., & Choi, S. Y. (2005). Tat-mediated protein transduction of human brain pyridoxal kinase into PC12 cells. Biochimie, 87(5), 481-487. https://doi.org/10.1016/j.biochi.2004.12.004

@article{78569b32d16f4f35a418ee3669ec7a50,

title = "Tat-mediated protein transduction of human brain pyridoxal kinase into PC12 cells",

abstract = "Pyridoxal kinase (PK) catalyses the phosphorylation of vitamin B 6 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.",

keywords = "Brain pyridoxal kinase, HIV-1 Tat, Protein therapy, Pyridoxal-5′-phosphate, Transduction",

author = "Kim, {Dae Won} and Kim, {Chung Kwon} and Choi, {Soo Hyun} and Choi, {Hee Soon} and Kim, {So Young} and An, {Jae Jin} and Lee, {Seung Ree} and Lee, {Sun Hwa} and Kwon, {Oh Shin} and Kang, {Tae Cheon} and Won, {Moo Ho} and Cho, {Yong Joon} and Cho, {Sung Woo} and Kang, {Jung Hoon} and Kim, {Tae Yoon} and Lee, {Kil Soo} and Jinseu Park and Eum, {Won Sik} and Choi, {Soo Young}",

year = "2005",

month = may,

doi = "10.1016/j.biochi.2004.12.004",

language = "English",

volume = "87",

pages = "481--487",

journal = "Biochimie",

issn = "0300-9084",

publisher = "Elsevier B.V.",

number = "5",

}

TY - JOUR

T1 - Tat-mediated protein transduction of human brain pyridoxal kinase into PC12 cells

AU - Kim, Dae Won

AU - Kim, Chung Kwon

AU - Choi, Soo Hyun

AU - Choi, Hee Soon

AU - Kim, So Young

AU - An, Jae Jin

AU - Lee, Seung Ree

AU - Lee, Sun Hwa

AU - Kwon, Oh Shin

AU - Kang, Tae Cheon

AU - Won, Moo Ho

AU - Cho, Yong Joon

AU - Cho, Sung Woo

AU - Kang, Jung Hoon

AU - Kim, Tae Yoon

AU - Lee, Kil Soo

AU - Park, Jinseu

AU - Eum, Won Sik

AU - Choi, Soo Young

PY - 2005/5

Y1 - 2005/5

N2 - Pyridoxal kinase (PK) catalyses the phosphorylation of vitamin B 6 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.

AB - Pyridoxal kinase (PK) catalyses the phosphorylation of vitamin B 6 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.

KW - Brain pyridoxal kinase

KW - HIV-1 Tat

KW - Protein therapy

KW - Pyridoxal-5′-phosphate

KW - Transduction

UR - http://www.scopus.com/inward/record.url?scp=20144387459&partnerID=8YFLogxK

U2 - 10.1016/j.biochi.2004.12.004

DO - 10.1016/j.biochi.2004.12.004

M3 - Article

C2 - 15820755

AN - SCOPUS:20144387459

SN - 0300-9084

VL - 87

SP - 481

EP - 487

JO - Biochimie

JF - Biochimie

IS - 5

ER -

Tat-mediated protein transduction of human brain pyridoxal kinase into PC12 cells

Abstract

Keywords

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this