Mechanism of the calcium-dependent self-association of bovine prothrombin. Use of a covalent cross-linking reagent to study the reaction.
The present study has made use of a covalent cross-linking agent, dithiobis(succinimidylpropionate), to study the self-association of prothrombin and has demonstrated that the covalent dimerization reaction involves the gamma-carboxyglutamic acid region of prothrombin (1-42 of 582). An essential role for the gamma-carboxyglutamic acid residues of prothrombin in the association reaction was demonstrated by experiments that converted gamma-carboxyglutamic acid residues to gamma-methylene glutamic acid or glutamic acid and resulted in a prothrombin species that was inactive in our cross-linking assay. Other experiments showed that very high concentrations of calcium ion inhibit the cross-linkage of prothrombin. This result is most consistent with an essential gamma-carboxyglutamic acid-calcium ion-gamma-carboxyglutamic acid bridge(s) in the calcium-dependent self-associated form of prothrombin.
D015055:1-Carboxyglutamic Acid; D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology*; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D002918:Chymotrypsin / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005973:Glutamine / Q000378:metabolism; D046911:Macromolecular Substances; D010446:Peptide Fragments / Q000378:metabolism; D011516:Prothrombin / Q000378:metabolism; D012964:Sodium / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013388:Succinimides / Q000494:pharmacology*
D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D010446:Peptide Fragments; D013388:Succinimides; D005973:Glutamine; D015055:1-Carboxyglutamic Acid; D011516:Prothrombin; D012964:Sodium; D002918:Chymotrypsin; C011240:dithiobis(succinimidylpropionate); D002118:Calcium