Linked Life Data

1918077

Source:http://linkedlifedata.com/resource/pubmed/id/1918077

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
29
pubmed:dateCreated
1991-11-14
pubmed:databankReference
pubmed:abstractText
An Mr 57,000 single-chain chimeric plasminogen activator, K12G0S32, consisting of a variable region fragment (Fv) derived from the fibrin fragment D-dimer-specific monoclonal antibody MA-15C5 and of a 33-kDa (amino acids Ala132 to Leu411) recombinant single-chain urokinase-type plasminogen activator (rscu-PA-33k) was studied. K12G0S32, secreted by infected Spodoptera frugiperda insect cells at a rate of 1.5 micrograms/10(6) cells/48 h, was purified to homogeneity by ion-exchange chromatography and gel filtration. It was obtained essentially as a single-chain molecule with a Ka = 5.5 x 10(9) M-1 for immobilized fragment D-dimer, similar to that of MA-15C5. The specific activity of both its single-chain and two-chain forms on fibrin plates was 100,000 IU/mg of urokinase-type plasminogen activator (u-PA) equivalent. Activation of plasminogen by two-chain K12G0S32 obeyed Michaelis-Menten kinetics with Km = 2.9 +/- 0.6 microM and a k2 = 3.7 +/- 0.6 s-1 (mean +/- S.D.; n = 3), as compared to Km = 12 microM and k2 = 4.8 s-1 for rtcu-PA-32k (recombinant low Mr two-chain u-PA consisting of amino acids Leu144 to Leu411). Single-chain K12G0S32 induced a dose- and time-dependent lysis of a 125I-fibrin-labeled human plasma clot immersed in citrated human plasma; 50% lysis in 2 h was obtained with 0.70 +/- 0.07 micrograms/ml (mean +/- S.D.; n = 5), as compared with 8.8 +/- 0.1 micrograms/ml for rscu-PA-32k (recombinant low Mr single-chain u-PA consisting of amino acids Leu144 to Leu411) (mean +/- S.D.; n = 3). With two-chain K12G0S32, 50% clot lysis in 2 h required 0.25 +/- 0.03 micrograms/ml (mean +/- S.D.; n = 3), as compared with only 0.62 +/- 0.04 micrograms/ml (mean +/- S.D.; n = 2) for rtcu-PA-32k. These results indicate that low Mr single-chain u-PA can be targeted to a fibrin clot with a single-chain Fv fragment of a fibrin-specific antibody, resulting in a 13-fold increase of the fibrinolytic potency of the single-chain form and a 2.5-fold increase of the potency of the two-chain form.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
266
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
19717-24
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:1918077-Amides, pubmed-meshheading:1918077-Amino Acid Sequence, pubmed-meshheading:1918077-Animals, pubmed-meshheading:1918077-Antibodies, Monoclonal, pubmed-meshheading:1918077-Base Sequence, pubmed-meshheading:1918077-Chimera, pubmed-meshheading:1918077-Cricetinae, pubmed-meshheading:1918077-DNA, pubmed-meshheading:1918077-Dithioerythritol, pubmed-meshheading:1918077-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:1918077-Fibrin, pubmed-meshheading:1918077-Fibrinolysis, pubmed-meshheading:1918077-Gene Expression, pubmed-meshheading:1918077-Models, Molecular, pubmed-meshheading:1918077-Molecular Sequence Data, pubmed-meshheading:1918077-Moths, pubmed-meshheading:1918077-Mutagenesis, Site-Directed, pubmed-meshheading:1918077-Oxidation-Reduction, pubmed-meshheading:1918077-Plasminogen Activators, pubmed-meshheading:1918077-Urokinase-Type Plasminogen Activator
pubmed:year
1991
pubmed:articleTitle
Characterization of a chimeric plasminogen activator consisting of a single-chain Fv fragment derived from a fibrin fragment D-dimer-specific antibody and a truncated single-chain urokinase.
pubmed:affiliation
Center for Thrombosis and Vascular Research, University of Leuven, Belgium.
pubmed:publicationType
Journal Article