Phosphorylation of purified recombinant hepatitis B virus-X protein by mitogen-activated protein kinase and protein kinase C in vitro

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Title
Phosphorylation of purified recombinant hepatitis B virus-X protein by mitogen-activated protein kinase and protein kinase C in vitro
Author(s)
Young Ik Lee; Sun Ok Kim; Hyok Joon Kwon; Jong Gu Park; Mi Jin Sohn; Soon Seok Jeong
Bibliographic Citation
Journal of Virological Methods, vol. 95, no. 1, pp. 1-10
Publication Year
2001
Abstract
The recombinant human hepatitis B virus-X protein (rhHBx) has been expressed as inclusion bodies in Escherichia coli and purified. By sequential dialysis of urea, rhHBx was folded into the native structure, which was demonstrated by both the efficacy of its transcriptional activation of the adenovirus major late promoter, fluorescence and circular dichroism (CD) analysis. The increase in CD values at 220 nm and a corresponding blue shift of the intrinsic fluorescence emission confirmed the ability of HBx to refold in lower concentrations of urea to produce the active protein. After purification and renaturation, the rhHBx protein was found to be phosphorylated by protein kinase C (PKC) and mitogen-activated protein kinase (MAPK). In vivo phosphorylation of HBx was also demonstrated. Although PKC and MAPK enhance the HBx phosphorylation in vitro, neither protein kinase A nor caseine kinase II (CKII) phosphorylate HBx protein, though there are possible substrate residues of both kinases in HBx protein. Phosphoamino acid analysis of the total acid hydrolyzed HBx showed that serine residues can be phosphorylated by PKC or MAPK.
Keyword
hepatitis B virus-X proteincircular dichroismprotein kinase Cmitogen-activated protein kinase
ISSN
0166-0934
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/S0166-0934(00)00282-2
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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