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1 THE EFFECT OF GLYCOSAMINOGLYCANS UPON THE FORMATION OF COMPLEXES BETWEEN FACTOR β-XIIa AND ANTITHROMBIN III
  1. A. S. Brecher,
  2. A. L. Spayde
  1. Bowling Green, OH

Abstract

This investigation explored the effect of glycosaminoglycans upon the initial enzyme in the intrinsic blood coagulation pathway, namely, Factor β--XIIa, and its interaction with antithrombin III (ATIII). Primary and secondary complexes of β--XIIa with ATIII were observed by SDS-PAGE and Western blot analysis utilizing a polyclonal human ATIII antibody to identify the complexes which were subsequently quantitated. With 3.15 μg β-XIIa: 3.15 μg ATIII pre-incubations for 15 min at RT, it was noted that 7.4 and 1.7% of the total protein in the SDS-PAGE lane was present as primary and secondary complexes, respectively, where the 2° complex is believed to be a proteolytic degradation product of the 1° complex. In the presence of a prior ATIII/12.6 μg heparin pre-incubation, the 1° complex increased to 34.1% of the protein in the lane, while the 2° complex increased to 12.1%. Upon replacing heparin with dermatan sulfate (chondroitin sulfate B), an increase of the 1° complex to 19.2% was noted whereas the 2° complex increased to 4.8%. Chondroitin sulfates A and C, by and large, exhibited no major changes in the 1° and 2° complex formation. These data indicate that both heparin and dermatan sulfate exhibit anticoagulant effects in the intrinsic blood clotting system and suggest that disorders in glycosaminoglycan production and degradation may be factors to consider in coagulopathies.

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