| pubmed-article:1751488 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:1751488 | lifeskim:mentions | umls-concept:C0086418 | lld:lifeskim |
| pubmed-article:1751488 | lifeskim:mentions | umls-concept:C0014834 | lld:lifeskim |
| pubmed-article:1751488 | lifeskim:mentions | umls-concept:C1171362 | lld:lifeskim |
| pubmed-article:1751488 | lifeskim:mentions | umls-concept:C0017262 | lld:lifeskim |
| pubmed-article:1751488 | lifeskim:mentions | umls-concept:C1515670 | lld:lifeskim |
| pubmed-article:1751488 | lifeskim:mentions | umls-concept:C1305923 | lld:lifeskim |
| pubmed-article:1751488 | lifeskim:mentions | umls-concept:C1709915 | lld:lifeskim |
| pubmed-article:1751488 | lifeskim:mentions | umls-concept:C0205195 | lld:lifeskim |
| pubmed-article:1751488 | pubmed:issue | 50 | lld:pubmed |
| pubmed-article:1751488 | pubmed:dateCreated | 1992-1-30 | lld:pubmed |
| pubmed-article:1751488 | pubmed:abstractText | Human zeta-thrombin, a catalytically competent serine proteinase, arises from a single chymotryptic cleavage at Trp-148 in alpha-thrombin to generate two nonconvalently associated polypeptide segments designated zeta 1-thrombin (the 36-residue A-chain disulfide linked to B-chain residues B1-148) and zeta 2-thrombin (B149-259). We report here the expression of recombinant zeta 2-thrombin in Escherichia coli and the reconstitution of catalytically competent zeta-thrombin by combination of zeta 1-thrombin with recombinant zeta 2-thrombin. A DNA fragment encoding zeta 2-thrombin was cloned into a pATH2 expression vector as a trpE-zeta 2 fusion gene, in which a factor Xa cleavage site was inserted between the trpE and the zeta 2-thrombin gene. High-level expression of this fusion protein was achieved under the control of the E. coli trp promoter. The expressed zeta 2-thrombin was liberated from the fusion protein by factor Xa cleavage, reduced with DTT, and purified to homogeneity by reverse-phase HPLC. Oxidation of the reduced zeta 2-thrombin in the presence of 80 microM CuSO4 and 6 M urea at pH 8.15 yielded material that was indistinguishable on HPLC from zeta 2-thrombin isolated by resolution of human zeta-thrombin. Catalytically active zeta-thrombin was generated by combination of recombinant zeta 2-thrombin with zeta 1-thrombin that was isolated by resolution of human zeta-thrombin. Recombinant zeta-thrombin displayed catalytic activities, toward a small chromogenic substrate and fibrinogen, that were similar to those of alpha-thrombin prepared from human blood plasma and zeta-thrombin obtained by treatment of alpha-thrombin with chymotrypsin.(ABSTRACT TRUNCATED AT 250 WORDS) | lld:pubmed |
| pubmed-article:1751488 | pubmed:language | eng | lld:pubmed |
| pubmed-article:1751488 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1751488 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:1751488 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1751488 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1751488 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1751488 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:1751488 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:1751488 | pubmed:month | Dec | lld:pubmed |
| pubmed-article:1751488 | pubmed:issn | 0006-2960 | lld:pubmed |
| pubmed-article:1751488 | pubmed:author | pubmed-author:TanE TET | lld:pubmed |
| pubmed-article:1751488 | pubmed:author | pubmed-author:ShaferJ AJA | lld:pubmed |
| pubmed-article:1751488 | pubmed:author | pubmed-author:LewisS DSD | lld:pubmed |
| pubmed-article:1751488 | pubmed:author | pubmed-author:StoneJ RJR | lld:pubmed |
| pubmed-article:1751488 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:1751488 | pubmed:day | 17 | lld:pubmed |
| pubmed-article:1751488 | pubmed:volume | 30 | lld:pubmed |
| pubmed-article:1751488 | pubmed:geneSymbol | trpE | lld:pubmed |
| pubmed-article:1751488 | pubmed:geneSymbol | trp | lld:pubmed |
| pubmed-article:1751488 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:1751488 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:1751488 | pubmed:pagination | 11694-9 | lld:pubmed |
| pubmed-article:1751488 | pubmed:dateRevised | 2008-11-21 | lld:pubmed |
| pubmed-article:1751488 | pubmed:meshHeading | pubmed-meshheading:1751488-... | lld:pubmed |
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| pubmed-article:1751488 | pubmed:meshHeading | pubmed-meshheading:1751488-... | lld:pubmed |
| pubmed-article:1751488 | pubmed:meshHeading | pubmed-meshheading:1751488-... | lld:pubmed |
| pubmed-article:1751488 | pubmed:meshHeading | pubmed-meshheading:1751488-... | lld:pubmed |
| pubmed-article:1751488 | pubmed:meshHeading | pubmed-meshheading:1751488-... | lld:pubmed |
| pubmed-article:1751488 | pubmed:meshHeading | pubmed-meshheading:1751488-... | lld:pubmed |
| pubmed-article:1751488 | pubmed:year | 1991 | lld:pubmed |
| pubmed-article:1751488 | pubmed:articleTitle | Reconstitution of catalytically competent human zeta-thrombin by combination of zeta-thrombin residues A1-36 and B1-148 and an Escherichia coli expressed polypeptide corresponding to zeta-thrombin residues B149-259. | lld:pubmed |
| pubmed-article:1751488 | pubmed:affiliation | Department of Biological Chemistry, Merck Sharp & Dohme Research Laboratories, West Point, Pennsylvania 19486. | lld:pubmed |
| pubmed-article:1751488 | pubmed:publicationType | Journal Article | lld:pubmed |
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