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Prokaryotic expression, purification and characterization of human cyclooxygenase-2

  • Authors:
    • Xiangzhi Liao
    • Wenhan Wang
    • Chuanxi Fan
    • Ning Yang
    • Jialiang Zhao
    • Ying Zhang
    • Ruijuan Gao
    • Guannan Shen
    • Simin Xia
    • Guiying Li
  • View Affiliations / Copyright

    Affiliations: Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, College of Life Sciences, Jilin University, Changchun, Jilin 130012, P.R. China
    Copyright: © Liao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 75-82
    |
    Published online on: May 31, 2017
       https://doi.org/10.3892/ijmm.2017.3007
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Abstract

Cyclooxygenase-2 (COX-2) is a key enzyme which catalyzes the conversion of arachidonic acid (AA) into prostaglandins (PGs). It plays an important role in pathophysiological processes, such as tumorigenesis, angiogenesis, inflammation and tumor cell drug resistance. Therefore, COX-2 has been viewed as an important target for cancer therapy. The preparation of COX-2 protein is an important initial step for the subsequent development of COX-2 inhibitors. In this study, we report a strategy to heterologously express truncated human COX-2 (trCOX-2) in Escherichia coli (E. coli) BL21(DE3) host cells. Following denaturation, purification and renaturation, we successfully obtained enzymatically active trCOX-2 containing 257 residues of the C-terminus. Homology modeling and molecular docking analyses revealed that trCOX-2 retained the predicted 3D catalytic domain structure and AA could still bind to its hydrophobic groove. Western blot analysis and ELISA indicated that the trCOX-2 still retained its characteristic antigenicity and binding activity, while COX assays revealed that trCOX-2 maintained its enzyme activity. On the whole, in this study, we provided a novel method to isolate trCOX-2 possessing AA binding and catalytic activities. This study thus lays a foundation to facilitate further investigations of COX-2 and offers a valuable method with which to achieve the prokaryotic expression of a eukaryotic membrane protein.
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Copy and paste a formatted citation
Spandidos Publications style
Liao X, Wang W, Fan C, Yang N, Zhao J, Zhang Y, Gao R, Shen G, Xia S, Li G, Li G, et al: Prokaryotic expression, purification and characterization of human cyclooxygenase-2. Int J Mol Med 40: 75-82, 2017.
APA
Liao, X., Wang, W., Fan, C., Yang, N., Zhao, J., Zhang, Y. ... Li, G. (2017). Prokaryotic expression, purification and characterization of human cyclooxygenase-2. International Journal of Molecular Medicine, 40, 75-82. https://doi.org/10.3892/ijmm.2017.3007
MLA
Liao, X., Wang, W., Fan, C., Yang, N., Zhao, J., Zhang, Y., Gao, R., Shen, G., Xia, S., Li, G."Prokaryotic expression, purification and characterization of human cyclooxygenase-2". International Journal of Molecular Medicine 40.1 (2017): 75-82.
Chicago
Liao, X., Wang, W., Fan, C., Yang, N., Zhao, J., Zhang, Y., Gao, R., Shen, G., Xia, S., Li, G."Prokaryotic expression, purification and characterization of human cyclooxygenase-2". International Journal of Molecular Medicine 40, no. 1 (2017): 75-82. https://doi.org/10.3892/ijmm.2017.3007
Copy and paste a formatted citation
x
Spandidos Publications style
Liao X, Wang W, Fan C, Yang N, Zhao J, Zhang Y, Gao R, Shen G, Xia S, Li G, Li G, et al: Prokaryotic expression, purification and characterization of human cyclooxygenase-2. Int J Mol Med 40: 75-82, 2017.
APA
Liao, X., Wang, W., Fan, C., Yang, N., Zhao, J., Zhang, Y. ... Li, G. (2017). Prokaryotic expression, purification and characterization of human cyclooxygenase-2. International Journal of Molecular Medicine, 40, 75-82. https://doi.org/10.3892/ijmm.2017.3007
MLA
Liao, X., Wang, W., Fan, C., Yang, N., Zhao, J., Zhang, Y., Gao, R., Shen, G., Xia, S., Li, G."Prokaryotic expression, purification and characterization of human cyclooxygenase-2". International Journal of Molecular Medicine 40.1 (2017): 75-82.
Chicago
Liao, X., Wang, W., Fan, C., Yang, N., Zhao, J., Zhang, Y., Gao, R., Shen, G., Xia, S., Li, G."Prokaryotic expression, purification and characterization of human cyclooxygenase-2". International Journal of Molecular Medicine 40, no. 1 (2017): 75-82. https://doi.org/10.3892/ijmm.2017.3007
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