Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
|
| pubmed:dateCreated |
1991-10-15
|
| pubmed:abstractText |
A cDNA construct (approximately 1 kb) of human BM-40 in a plasmid with the cytomegalovirus promoter and enhancer was used to produce several stable clones by transfecting two human cell lines (293, HT 1080). These clones showed a high expression of exogenous 1-kb BM-40 mRNA and no or only little endogenous 2.2-kb mRNA. These clones also secreted BM-40 at high rates (5-50 micrograms ml-1 day-1) into serum-free culture medium as shown by electrophoresis, radioimmunoassay and metabolic labelling. Transfection with the plasmid and overexpression of BM-40 had no effect on cell spreading, proliferation rate and adhesion patterns to extracellular matrix substrates. Recombinant human BM-40 was purified by anion-exchange chromatography and showed the expected N-terminal sequence and amino acid composition. The protein was also identical or similar to authentic BM-40 purified from the mouse Engelbreth-Holm-Swarm tumor in hexosamine content, electrophoretic mobility, circular dichroism and binding activity for calcium and collagen IV. Reduction of both authentic and recombinant BM-40 decreased binding activity which indicates correct formation of disulfide bonds in the recombinant protein. A specific and sensitive radioimmunoassay for human BM-40 was shown to be useful for detecting small quantities of the protein in human cell culture medium and blood. No significant cross-reaction was, however, detected between human and mouse BM-40.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Collagen,
http://linkedlifedata.com/resource/pubmed/chemical/DNA,
http://linkedlifedata.com/resource/pubmed/chemical/Methionine,
http://linkedlifedata.com/resource/pubmed/chemical/Osteonectin,
http://linkedlifedata.com/resource/pubmed/chemical/RNA, Messenger,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Sep
|
| pubmed:issn |
0014-2956
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
1
|
| pubmed:volume |
200
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
529-36
|
| pubmed:dateRevised |
2008-11-21
|
| pubmed:meshHeading |
pubmed-meshheading:1653704-Amino Acid Sequence,
pubmed-meshheading:1653704-Animals,
pubmed-meshheading:1653704-Blotting, Northern,
pubmed-meshheading:1653704-Collagen,
pubmed-meshheading:1653704-Cytomegalovirus,
pubmed-meshheading:1653704-DNA,
pubmed-meshheading:1653704-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:1653704-Enhancer Elements, Genetic,
pubmed-meshheading:1653704-Gene Expression,
pubmed-meshheading:1653704-Genes, Viral,
pubmed-meshheading:1653704-Humans,
pubmed-meshheading:1653704-Methionine,
pubmed-meshheading:1653704-Mice,
pubmed-meshheading:1653704-Molecular Sequence Data,
pubmed-meshheading:1653704-Nucleic Acid Hybridization,
pubmed-meshheading:1653704-Osteonectin,
pubmed-meshheading:1653704-Plasmids,
pubmed-meshheading:1653704-Promoter Regions, Genetic,
pubmed-meshheading:1653704-RNA, Messenger,
pubmed-meshheading:1653704-Radioimmunoassay,
pubmed-meshheading:1653704-Recombinant Proteins,
pubmed-meshheading:1653704-Transfection
|
| pubmed:year |
1991
|
| pubmed:articleTitle |
Recombinant expression and properties of the human calcium-binding extracellular matrix protein BM-40.
|
| pubmed:affiliation |
Department of Dermatology, University of Munich, Federal Republic of Germany.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|