Expression, purification and molecular modeling of the NIa protease of Cardamom mosaic virus.
The NIa protease of Potyviridae is the major viral protease that processes potyviral polyproteins. The NIa protease coding region of Cardamom mosaic virus (CdMV) is amplified from the viral cDNA, cloned and expressed in Escherichia coli. NIa protease forms inclusion bodies in E.coli. The inclusion bodies are solubilized with 8 M urea, refolded and purified by Nickel-Nitrilotriacetic acid affinity chromatography. Three-dimensional modeling of the CdMV NIa protease is achieved by threading approach using the homologous X-ray crystallographic structure of Tobacco etch mosaic virus NIa protease. The model gave an insight in to the substrate specificities of the NIa proteases and predicted the complementation of nearby residues in the catalytic triad (H42, D74 and C141) mutants in the cis protease activity of CdMV NIa protease.
School of Biological Sciences, Madurai Kamaraj University, Madurai, 625021, Tamil Nadu, India. jebasingh@dbtiplsmku.org
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Jebasingh
Eswari P J
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Pandaranayaka
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Mahalakshmi
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Kasin Yadunandam
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Krishnaswamy
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R
Usha
D000595:Amino Acid Sequence; D018076:DNA, Complementary / Q000737:chemistry / Q000378:metabolism; D004279:DNA, Viral / Q000737:chemistry / Q000378:metabolism; D028523:Elettaria / Q000821:virology; D010450:Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000201:enzymology* / Q000378:metabolism; D013379:Substrate Specificity; D014764:Viral Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism
D018076:DNA, Complementary; D004279:DNA, Viral; D014764:Viral Proteins; D010450:Endopeptidases; C059992:nuclear inclusion protein a, mosaic viruses