상세 정보

underline
Metadata Downloads : dc(xml) or Excel
Cited 0 time in scopus ci

Title 

In vitro folding of recombinant hepatitis B virus X-protein produced in Escherichia coli: formation of folding intermediates

Authors 

Sun Ok KimMi Jin SohnSoon Seog JeongJeh Hoon ShinYoung Ik Lee

Publisher 

Springer Verlag (Germany)

Issue Date 

1999

Citation 

Journal of Biochemistry and Molecular Biology, vol. 32, no. 6, pp. 521-528

Keywords 

Circular dichroismFluorescence emission maximumHepatitis B virus X-proteinRefoldingReverse phase-high performance liquid chromatography

Abstract 

The folding of recombinant hepatitis B virus X-protein (rHBx) solubilized from Escherichia coli inclusion bodies was investigated. By sequential dialysis of urea, rHBx was folded into its native structure, which was demonstrated by the efficacy of its transcriptional activation of the adenovirus major late promoter (MLP), fluorescence spectroscopy, and circular dichroism (CD) analysis. The decrease in CD values at 220 nm and a corresponding blue shift of the intrinsic fluorescence emission confirmed the ability of rHBx to refold in lower concentrations of urea, yielding the active protein. Equilibrium and kinetic studies of the refolding of rHBx were carried out by tryptophan fluorescence measurements. From the biphasic nature of the fluorescence curves, the existence of stable intermediate states in the renaturation process was inferred. Reverse phase-high performance liquid chromatography (RP-HPLC) analysis further demonstrated the existence of these intermediates and their apparent compactness.

ISSN 

1225-8687

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


There are no files associated with this item.
qrcode

FusionCharts.
DSpace Software Coptright(c) 2010 MIT and Hewleft-Packard  /  KRIBB-REPOSITORY ( Email:jakim@kribb.re.kr)