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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
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pubmed:dateCreated |
1994-12-21
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pubmed:abstractText |
We have expressed recombinant human apolipoprotein A-II (apoA-II) in Escherichia coli, as a fusion protein with Schistosoma japonicum glutathione-S-transferase (GST). The GST-AII fusion protein was recovered by affinity chromatography using glutathione as a ligand. After thrombin cleavage and removal of the GST carrier, recombinant apoA-II was obtained in a highly purified form and was exclusively composed of dimeric apoA-II. Kinetics of association to dimyristoylglycerophosphocholine (Myr2GroPCho) vesicles showed that recombinant apoA-II exhibited the same pattern of association as human plasma apoA-II. Electron microscopic analysis of the complexes showed a typical pattern of rouleaux, characteristic of stacked discs, with a diameter similar to that determined by gradient-gel electrophoresis. Circular dichroism measurements showed that the alpha-helical content of both plasma and recombinant apoA-II increased similarly when the proteins associated with Myr2GroPCho vesicles, at the expense of a random-coil structure. Lipid-bound apoA-II consisted of 70-72% alpha helices, suggesting the presence of three 18-residue alpha helices/apoA-II monomer. Cross-linking experiments indicated that Myr2GroPCho complexes contained two molecules dimeric apoA-II/vesicle. Recombinant apoA-II was as efficient as plasma apoA-II in associating with HDL subclasses, and in displacing apoA-I from dipalmitoylglycerophosphocholine/cholesterol/apoA-I complexes, most likely due to its highly ordered secondary structure when associated with Myr2GroPCho vesicles. These findings demonstrate that recombinant apoA-II exhibits the same structural and functional properties as human plasma apoA-II. Thus, the expression system utilized is appropriate to produce mutagenized forms to further structure/function analysis.
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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/Apolipoprotein A-I,
http://linkedlifedata.com/resource/pubmed/chemical/Apolipoprotein A-II,
http://linkedlifedata.com/resource/pubmed/chemical/Dimyristoylphosphatidylcholine,
http://linkedlifedata.com/resource/pubmed/chemical/Glutathione Transferase,
http://linkedlifedata.com/resource/pubmed/chemical/Lipoproteins, HDL,
http://linkedlifedata.com/resource/pubmed/chemical/Liposomes,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Fusion Proteins
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pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
0014-2956
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
225
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1141-50
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pubmed:dateRevised |
2007-7-23
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pubmed:meshHeading |
pubmed-meshheading:7957205-Amino Acid Sequence,
pubmed-meshheading:7957205-Animals,
pubmed-meshheading:7957205-Apolipoprotein A-I,
pubmed-meshheading:7957205-Apolipoprotein A-II,
pubmed-meshheading:7957205-Binding, Competitive,
pubmed-meshheading:7957205-Dimyristoylphosphatidylcholine,
pubmed-meshheading:7957205-Drug Stability,
pubmed-meshheading:7957205-Escherichia coli,
pubmed-meshheading:7957205-Gene Expression,
pubmed-meshheading:7957205-Glutathione Transferase,
pubmed-meshheading:7957205-Humans,
pubmed-meshheading:7957205-Lipoproteins, HDL,
pubmed-meshheading:7957205-Liposomes,
pubmed-meshheading:7957205-Molecular Sequence Data,
pubmed-meshheading:7957205-Recombinant Fusion Proteins,
pubmed-meshheading:7957205-Schistosoma japonicum
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pubmed:year |
1994
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pubmed:articleTitle |
Purification and characterization of recombinant human apolipoprotein A-II expressed in Escherichia coli.
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pubmed:affiliation |
URA CNRS 1283, Institut des Cordeliers, Paris, France.
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pubmed:publicationType |
Journal Article,
In Vitro,
Research Support, Non-U.S. Gov't
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