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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
9
|
| pubmed:dateCreated |
1987-5-6
|
| pubmed:abstractText |
We produced human apolipoprotein A-I (apoA-I) in Chinese hamster ovary (CHO) cells. The CHO cells were transfected with an expression plasmid which placed the human apoA-I gene under the direction of the human metallothionein II gene promoter. Isolation of a clonal cell line resulted in high level expression of apoA-I. Greater than 30% of total protein secreted by these CHO cells was apoA-I, which enabled us to purify apoA-I with a single step purification scheme. As a result, large quantities of apoA-I can be produced and isolated without having to rely on plasma sources. Structural characterization of the recombinant apoA-I showed it to be identical to authentic apoA-I from human serum high density lipoprotein. Furthermore, we demonstrated approximately equal to 90% of the apoA-I secreted by CHO cells is processed, mature protein. A portion of the secreted recombinant apoA-I was associated with lipid and floated at a density approximately equal to 1.10 g/ml. Additional analysis identified the presence of five isoforms of apoA-I in the CHO cell conditioned medium. Processing and post-translational modification of the recombinant apoA-I occurred in the CHO cell cultures in the absence of serum components. We conclude that the human apoA-I produced by CHO cells is identical to circulating, mature apoA-I in humans and that recombinant mammalian expression offers an opportunity to investigate apoA-I processing.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Apolipoprotein A-I,
http://linkedlifedata.com/resource/pubmed/chemical/Apolipoproteins A,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Recombinant,
http://linkedlifedata.com/resource/pubmed/chemical/Metallothionein,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Mar
|
| pubmed:issn |
0021-9258
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
25
|
| pubmed:volume |
262
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
4241-7
|
| pubmed:dateRevised |
2008-11-21
|
| pubmed:meshHeading |
pubmed-meshheading:3104338-Amino Acid Sequence,
pubmed-meshheading:3104338-Animals,
pubmed-meshheading:3104338-Apolipoprotein A-I,
pubmed-meshheading:3104338-Apolipoproteins A,
pubmed-meshheading:3104338-Cell Line,
pubmed-meshheading:3104338-Centrifugation, Density Gradient,
pubmed-meshheading:3104338-Clone Cells,
pubmed-meshheading:3104338-Cricetinae,
pubmed-meshheading:3104338-DNA, Recombinant,
pubmed-meshheading:3104338-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:3104338-Female,
pubmed-meshheading:3104338-Humans,
pubmed-meshheading:3104338-Metallothionein,
pubmed-meshheading:3104338-Ovary,
pubmed-meshheading:3104338-Plasmids,
pubmed-meshheading:3104338-Promoter Regions, Genetic,
pubmed-meshheading:3104338-Protein Processing, Post-Translational,
pubmed-meshheading:3104338-Recombinant Proteins,
pubmed-meshheading:3104338-Transfection
|
| pubmed:year |
1987
|
| pubmed:articleTitle |
Expression and characterization of human apolipoprotein A-I in Chinese hamster ovary cells.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|