Voprosy meditsinskoi khimii (ISSN 0042-8809)

Use of affinity chromatography for isolating prethrombin 1

   
Strukova S.M., Mitroshina T.N., Umarova V.A., Kudriashov B.A.
PubMed Id: 7336661
Year: 1981 vol: 27  issue:6  pages: 840-844
Abstract: A procedure is described for isolation of prethrombin I using biospecific chromatography of the thrombin hydrolysate of prothrombin complex on heparin-Sepharose. The prothrombin complex was activated in 0.05 M Tris-HCl buffer, pH 7.4, containing 0.15 M NaCl, ImM EDTA-Na salt at 37 degrees within 15-20 min; the ratio of the enzyme and substrate was equal to 1 : 1300 (evaluated by the enzymatic activity). Then prethrombin I was isolated by the affinity chromatography. Prethrombin I was specially adsorbed on the immobilized heparin and eluted with 0.45 M NaCl. The substance was free from factor X and alpha-thrombin, which were eluted with 0.64 M and 0.8-1.0 M NaCl, respectively. Fragment I of prothrombin was not adsorbed on heparin-Sepharose and was found in the free volume of column. Preparation of prethrombin I (molecular mass 67,000 +/- 3,000) did not exhibit the coagulating, esterase and prothrombin activities but produced thrombin in presence of factor Xa. The abilities of prethrombin I to interact with blood vessel chemoreceptors and to activate the anticoagulation system were shown, when physiological activity of prethrombin I was studied using perfusion of rabbit carotid sinus, which was insulated from humoral action but retained the nervous connections with a body.
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Reference: Strukova S.M., Mitroshina T.N., Umarova V.A., Kudriashov B.A., Use of affinity chromatography for isolating prethrombin 1, Voprosy meditsinskoi khimii, 1981, vol: 27(6), 840-844.
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