Source:http://linkedlifedata.com/resource/pubmed/id/11049742
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
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| pubmed:dateCreated |
2001-1-3
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| pubmed:abstractText |
Both glycosylated and nonglycosylated forms of recombinant human prourokinase were produced to the level of 20 mg/L by yeast Pichia pastoris in BMMY medium after 2 days of culture. The expressed pro-UK was 98% secreted into the culture medium and easily purified by carboxymethyl cellulose chromatography. More than 99% of pro-UK in the culture medium was found in single-chain form. This was contradictory to a previous finding which found that glycosylation of pro-UK by yeast inhibited its secretion. The absence of glycosylation at Asn302 of pro-UK has no measurable effect on its secretion from the yeast cells. However, the nonglycosylated pro-UK was much less stable in the culture medium, probably due to proteolysis. Nonglycosylated pro-UK from yeast had a clot lysing activity comparable to that of Escherichia coli-derived or mammalian cell-derived recombinant pro-UK. By contrast, the glycosylated yeast pro-UK was less activatable by plasmin and had a lower enzymatic activity against plasminogen and a lower clot lysing activity than nonglycosylated pro-UK from yeast, while their amidolytic activity against S2444 was equivalent. It was concluded that glycosylation of pro-UK by yeast P. pastoris interferes with the catalytic site but not secretion of this protein.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Carboxymethylcellulose Sodium,
http://linkedlifedata.com/resource/pubmed/chemical/Fibrinogen,
http://linkedlifedata.com/resource/pubmed/chemical/Fibrinolysin,
http://linkedlifedata.com/resource/pubmed/chemical/Plasminogen,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Urokinase-Type Plasminogen Activator,
http://linkedlifedata.com/resource/pubmed/chemical/saruplase
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| pubmed:status |
MEDLINE
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| pubmed:month |
Nov
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| pubmed:issn |
1046-5928
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| pubmed:author | |
| pubmed:copyrightInfo |
Copyright 2000 Academic Press.
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| pubmed:issnType |
Print
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| pubmed:volume |
20
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
179-85
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| pubmed:dateRevised |
2010-11-18
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| pubmed:meshHeading |
pubmed-meshheading:11049742-Binding Sites,
pubmed-meshheading:11049742-Carboxymethylcellulose Sodium,
pubmed-meshheading:11049742-Chromatography, Ion Exchange,
pubmed-meshheading:11049742-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:11049742-Enzyme Activation,
pubmed-meshheading:11049742-Enzyme Stability,
pubmed-meshheading:11049742-Fibrinogen,
pubmed-meshheading:11049742-Fibrinolysin,
pubmed-meshheading:11049742-Glycosylation,
pubmed-meshheading:11049742-Humans,
pubmed-meshheading:11049742-Kinetics,
pubmed-meshheading:11049742-Mutagenesis, Site-Directed,
pubmed-meshheading:11049742-Mutation,
pubmed-meshheading:11049742-Pichia,
pubmed-meshheading:11049742-Plasminogen,
pubmed-meshheading:11049742-Recombinant Proteins,
pubmed-meshheading:11049742-Urokinase-Type Plasminogen Activator
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| pubmed:year |
2000
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| pubmed:articleTitle |
Glycosylation of prourokinase produced by Pichia pastoris impairs enzymatic activity but not secretion.
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| pubmed:affiliation |
Institute of Molecular Medicine, Nanjing, 210093, People's Republic of China.
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| pubmed:publicationType |
Journal Article
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