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Mol. Cells 2006; 22(1): 13-20
Published online January 1, 1970
© The Korean Society for Molecular and Cellular Biology
Molecular and Structural Characterization of the Domain 2 of Hepatitis C Virus Non-structural Protein 5A
Hepatitis C virus (HCV) non-structural protein 5A protein (NS5A), which consists of three functional domains, is involved in regulating viral replication, interferon resistance, and apoptosis. Recently, the three-dimensional structure of the domain 1 was determined. However, currently the molecular basis for the domains 2 and 3 of HCV NS5A is yet to be defined. Toward this end, we expressed, purified the domain 2 of the NS5A (NS5A-D2), and then performed biochemical and structural studies. The purified domain 2 was active and was able to bind NS5B and PKR, biological partners of NS5A. The results from gel filtration, CD analysis, 1D 1H NMR and 2D 1H-15N heteronuclear single quantum correlation (HSQC) spectroscopy indicate that the domain 2 of NS5A appears to be flexible and disordered.
Keywords Expression and Purification; Hepatitis C Virus NS5A; NMR; NS5A Domain 2; NS5B.
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Research Article
Mol. Cells 2006; 22(1): 13-20
Published online August 31, 2006
Copyright © The Korean Society for Molecular and Cellular Biology.
Molecular and Structural Characterization of the Domain 2 of Hepatitis C Virus Non-structural Protein 5A
Yu Liang, Cong Bao Kang, Ho Sup Yoon
Abstract
Hepatitis C virus (HCV) non-structural protein 5A protein (NS5A), which consists of three functional domains, is involved in regulating viral replication, interferon resistance, and apoptosis. Recently, the three-dimensional structure of the domain 1 was determined. However, currently the molecular basis for the domains 2 and 3 of HCV NS5A is yet to be defined. Toward this end, we expressed, purified the domain 2 of the NS5A (NS5A-D2), and then performed biochemical and structural studies. The purified domain 2 was active and was able to bind NS5B and PKR, biological partners of NS5A. The results from gel filtration, CD analysis, 1D 1H NMR and 2D 1H-15N heteronuclear single quantum correlation (HSQC) spectroscopy indicate that the domain 2 of NS5A appears to be flexible and disordered.
Keywords: Expression and Purification, Hepatitis C Virus NS5A, NMR, NS5A Domain 2, NS5B.
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