Cited 5 times in

Cu/Zn Incorporation During Purification of Soluble Human EC-SOD from E. coli Stabilizes Proper Disulfide Bond Formation

Title
Cu/Zn Incorporation During Purification of Soluble Human EC-SOD from E. coli Stabilizes Proper Disulfide Bond Formation
Authors
Ji-Young Bae;Bon-Kyung Koo;Han Choe;Hyang Kyu Lee;Young-Jin Son;Thu Trang Thi Vu;Minh Tan Nguyen;Jung-A Song;Han-Bong Ryu
Issue Date
2013
Journal Title
Applied Biochemistry and Biotechnology
ISSN
0273-2289
Citation
Applied Biochemistry and Biotechnology, Vol.169(5) : 1633~1647, 2013
Abstract
Extracellular superoxide dismutase (EC-SOD) is the only enzyme that removes superoxide radical in the extracellular space. The reduction of EC-SOD is linked to many diseases, suggesting that the protein may have therapeutic value. EC-SOD is reported to be insoluble and to make inclusion bodies when overexpressed in the cytoplasm of Escherichia coli. The refolding process has the advantage of high yield, but has the disadvantage of frequent aggregation or misfolding during purification. For the first time, this study shows that fusion with maltose-binding protein (MBP), N-utilization substance protein A, and proteindisulfide isomerase enabled the soluble overexpression of EC-SOD in the cytoplasm of E. coli. MBP-tagged human EC-SOD(hEC-SOD) was purified by MBP affinity and anion exchange chromatography, and its identity was confirmed by MALDI-TOF MS analysis. The purified protein showed good enzyme activity in vitro; however, there was a difference in metal binding. When copper and zinc were incorporated into hEC-SOD before MBP tag cleavage, the enzymatic activity was higher than when the metal ions were bound to the purified protein after MBP tag cleavage. Therefore, the enzymatic activity of hEC-SOD is associated with metal incorporation and protein folding via disulfide bond.
URI
http://link.springer.com/article/10.1007%2Fs12010-012-0025-x

http://ir.ymlib.yonsei.ac.kr/handle/22282913/86642
DOI
 10.1007/s12010-012-0025-x.
Appears in Collections:
1. 연구논문 > 3. College of Nursing > Dept. of Nursing Environment Systems
Yonsei Authors
사서에게 알리기
  feedback
Files in This Item:
Export
RIS (EndNote)
XLS (Excel)
XML

qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse