Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
|
| pubmed:dateCreated |
1990-12-24
|
| pubmed:abstractText |
Rat surfactant protein A (SP-A) was expressed in a Chinese hamster ovary (CHO-K1) cell line and characterized for biologic activity using assays for receptor binding and modulation of phospholipid secretion from isolated type II cells. The CHO-K1 cell line was cotransfected with separate plasmids encoding for the rat SP-A, dihydrofolate reductase and neomycin phosphotransferase, respectively. Antibiotic (Geneticin-G418)-resistant transformants were screened by ELISA for the secretion of recombinant SP-A into the media. Northern analysis of the transfected cell lines demonstrated the expression of both 1.6 kb and 0.9 kb mRNA species for SP-A, consistent with the proposed differential polyadenylation of the primary transcript. Amplification with methotrexate resulted in a dose-dependent increase in mRNA for SP-A and a 20-fold increase in the production of recombinant SP-A relative to untreated cells. Maximum production of SP-A was 370 micrograms of SP-A/l of media in a 4-day incubation. Recombinant SP-A was purified from the serum-free media of large scale cultures of transfected, amplified CHO cells by affinity chromatography on mannose-Sepharose. The recombinant SP-A migrated similarly to native SP-A by NaDodSO4-PAGE analysis under reducing and nonreducing conditions and under reducing conditions after digestion with N-glycanase. Recombinant SP-A effectively competed with 125I-native SP-A for binding to the high affinity receptor for SP-A on isolated plasma membranes from rat alveolar type II cells. The recombinant SP-A was as effective as native SP-A in the inhibition of secretion of phospholipid from isolated type II cells. We conclude that recombinant rat SP-A produced in Chinese hamster ovary cells is physically and functionally similar to native rat SP-A.
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| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Gentamicins,
http://linkedlifedata.com/resource/pubmed/chemical/Kanamycin Kinase,
http://linkedlifedata.com/resource/pubmed/chemical/Methotrexate,
http://linkedlifedata.com/resource/pubmed/chemical/Phospholipids,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphotransferases,
http://linkedlifedata.com/resource/pubmed/chemical/Proteolipids,
http://linkedlifedata.com/resource/pubmed/chemical/Pulmonary Surfactant-Associated...,
http://linkedlifedata.com/resource/pubmed/chemical/Pulmonary Surfactant-Associated...,
http://linkedlifedata.com/resource/pubmed/chemical/Pulmonary Surfactants,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Tetrahydrofolate Dehydrogenase,
http://linkedlifedata.com/resource/pubmed/chemical/antibiotic G 418
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Oct
|
| pubmed:issn |
0006-3002
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
23
|
| pubmed:volume |
1087
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
190-8
|
| pubmed:dateRevised |
2007-11-14
|
| pubmed:meshHeading |
pubmed-meshheading:2171680-Animals,
pubmed-meshheading:2171680-Binding, Competitive,
pubmed-meshheading:2171680-Blotting, Northern,
pubmed-meshheading:2171680-Cell Line,
pubmed-meshheading:2171680-Cricetinae,
pubmed-meshheading:2171680-Drug Resistance,
pubmed-meshheading:2171680-Gene Amplification,
pubmed-meshheading:2171680-Gene Expression,
pubmed-meshheading:2171680-Gentamicins,
pubmed-meshheading:2171680-Kanamycin Kinase,
pubmed-meshheading:2171680-Methotrexate,
pubmed-meshheading:2171680-Phospholipids,
pubmed-meshheading:2171680-Phosphotransferases,
pubmed-meshheading:2171680-Plasmids,
pubmed-meshheading:2171680-Proteolipids,
pubmed-meshheading:2171680-Pulmonary Surfactant-Associated Protein A,
pubmed-meshheading:2171680-Pulmonary Surfactant-Associated Proteins,
pubmed-meshheading:2171680-Pulmonary Surfactants,
pubmed-meshheading:2171680-Rats,
pubmed-meshheading:2171680-Recombinant Proteins,
pubmed-meshheading:2171680-Tetrahydrofolate Dehydrogenase,
pubmed-meshheading:2171680-Transfection,
pubmed-meshheading:2171680-Transformation, Genetic
|
| pubmed:year |
1990
|
| pubmed:articleTitle |
Expression and characterization of rat surfactant protein A synthesized in Chinese hamster ovary cells.
|
| pubmed:affiliation |
Lord and Taylor Laboratory for Lung Biochemistry, Department of Medicine, National Jewish Center for Immunology and Respiratory Medicine, Denver, CO.
|
| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
|