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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
1
|
| pubmed:dateCreated |
1975-12-29
|
| pubmed:abstractText |
1. Monomeric nu-chains were conjugated with CNBr-activated Sepharose 4B. The C-terminal cysteine of the conjugated nu-chain was converted to a mixed disulfide with 3-carboxy-4-nitro-benzenethiol (Nbs) and used to separate plasma proteins with reactive thiol groups. The plasma proteins, alpha1-antitrypsin and prealbumin have the greatest affinity for the interchange reaction with mixed disulfides. The disulfide link between alpha1-antitrypsin and nu-chain is sensitive to excess Nbs, and is selectively cleaved in the presence of 5,5'-dithiobis(2-nitrobenzoate) (Nbs2) which accepts the sulfhydryl group of alpha1-antitrypsin. 2. A Simple method developed for the isolation of human alpha1-antitrypsin was equally effective for the various inherited phenotypes and for alpha1-antitrypsin from the dog, baboon, and monkey, Glutathione-Sepharose was also used successfully, but the nu-chain conjugate yielded alpha1-antitrypsin less contaminated with mercaptalbumin and prealbumin. 3. The alpha1-antitrypsin is harvested from this procedure as a mixed disulfide with Nbs. The negative charge of Nbs at pH 8.1 causes an increased electrophoretic mobility of the alpha1-antitrypsin derivative. Mild reduction liberates Nbs and electrophoretic mobility of alpha1-antitrypsin returns to normal. The method described can increase the alpha1-antitrypsin content of a plasma fraction from 5% of the total protein to 95% within one day with a yield of about 50%. This purification procedure does not exert any detectable effect on microheterogeneity.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Sep
|
| pubmed:issn |
0014-2956
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
1
|
| pubmed:volume |
57
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
107-13
|
| pubmed:dateRevised |
2007-7-23
|
| pubmed:meshHeading |
pubmed-meshheading:809280-Animals,
pubmed-meshheading:809280-Binding Sites,
pubmed-meshheading:809280-Chromatography, Affinity,
pubmed-meshheading:809280-Disulfides,
pubmed-meshheading:809280-Dogs,
pubmed-meshheading:809280-Haplorhini,
pubmed-meshheading:809280-Humans,
pubmed-meshheading:809280-Macromolecular Substances,
pubmed-meshheading:809280-Papio,
pubmed-meshheading:809280-Protein Binding,
pubmed-meshheading:809280-Spectrophotometry,
pubmed-meshheading:809280-Spectrophotometry, Ultraviolet,
pubmed-meshheading:809280-Sulfhydryl Compounds,
pubmed-meshheading:809280-alpha 1-Antitrypsin
|
| pubmed:year |
1975
|
| pubmed:articleTitle |
Purification of alpha1-antitrypsin from plasma through thiol-disulfide interchange.
|
| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
|