Influence of metal ions on prothrombin self-association. Demonstration of dimer formation by intermolecular cross-linking with dithiobis(succinimidylpropionate).

Abstract

Interaction of certain metal ions with prothrombin and prothrombin fragment 1 has been shown to result in conformational change(s). Self-association of prothrombin and prothrombin fragment 1 in the presence of divalent cations has been reported. The present study has made use of a covalent cross-linking reagent, dithiobis(succinimidylpropionate), to study the self-association of prothrombin in the presence of divalent cations. In the presence of certain divalent cations, prothrombin dimer is the product of such cross-linking. Optimal dimerization of bovine prothrombin requires preincubation with calcium ions prior to the addition of cross-linking reagent. Calcium ions are also required for dimerization after this preincubation period. A similar time dependence is not seen with human prothrombin. The dimerization of bovine prothrombin is also supported by strontium and gadolinium to an extent comparable to that of calcium and to a lesser extent by barium or manganese. Magnesium ions do not support dimerization. The results suggest that certain divalent cations either induce or stabilize a conformation of prothrombin which can self-associate to form dimers. The results further suggest that divalent cations are also necessary for the actual cross-linking process subsequent to this conformational change.