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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
2
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pubmed:dateCreated |
1998-2-18
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pubmed:abstractText |
Reported here is the production of recombinant human rhinovirus 14 (HRV14) 2A protease from bacterial cells transformed with a heat-inducible plasmid containing the HRV14 2A cDNA sequence. Overexpressed 2A protein partitioned into the inclusion bodies was solubilized in urea and then refolded in the presence of Zn2+ Transition metals were required for the restoration of 2A protease activity as a structural component, but appeared to be inhibitory if added exogenously once the enzyme was refolded. Based on the cleavage specificity studies, a colorimetric assay was developed for the highly purified HRV14 2A protease. A peptide with the sequence RKGDIKSY-p-nitroanilide was found to be cleaved by the 2A protease with a k(cat)/Km ratio of approximately 335 M(-1)s(-1), which allows its activity to be measured continuously with a spectrophotometer or a microplate reader.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-1331062,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-1649327,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-1915271,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-2160953,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-2252396,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-2831385,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-7515772,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-7595358,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-7941352,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-8230412,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-8338854,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-8380894,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-8392608,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-8396129,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-8520469,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-8535153,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-8580843,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-8761474,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-8861917,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-8873593,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-9207196,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9445078-9344409
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Feb
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pubmed:issn |
0022-538X
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
72
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1683-7
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
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pubmed:year |
1998
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pubmed:articleTitle |
Enzymatic characterization of refolded human rhinovirus type 14 2A protease expressed in Escherichia coli.
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pubmed:affiliation |
Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana 46285, USA. qmwang@lilly.com
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pubmed:publicationType |
Journal Article
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