The mechanism of activation of rabbit plasminogen by urokinase.
The data presented in this paper show that when rabbit plasminogen is activated to plasmin by urokinase at least two peptide bonds are cleaved in the process. Urokinase first cleaves an internal peptide bond in plasminogen, leading to two-chain disulfide-linked plasmin molecule. The plasmin heavy chain of molecular weight 66,000 to 69,000 possesses an NH2-terminal amino acid sequence identical with the original plasminogen (molecular weight 88,000 to 92,000). The plasmin light chain of molecular weight 24,000 to 26,000 is known to be derived from the COOH-terminal portion of plasminogen. The plasmin generated during the activation of plasminogen is capable, by a feedback process, of cleaving a peptide of molecular weight 6,000 to 8,000 from the NH2 terminus of the heavy chain, producing a proteolytically modified heavy chain of molecular weight 58,000 to 62,000. Plasmin also can cleave this same peptide from the original plasminogen, yielding an altered plasminogen of molecular weight 82,000 to 86,000. This plasmin-altered plasminogen and the plasmin heavy chain derived from it by urokinase activation process NH2-terminal amino acid sequences which are identical with each other and with the plasminolytic product of the original plasmin heavy chain. These studies support a mechanism of activation of plasminogen by urokinase which involves loss of a peptide located on the NH2 terminus of plasminogen. However, these same results show that this NH2-terminal peptide need not be released from rabbit plasminogen prior to the cleavage of the internal peptide bond which leads to the two-chain plasmin molecule. Furthermore, these studies show that urokinase cannot remove this peptide from either the original rabbit plasminogen molecule or from the heavy chain of the initial plasmin formed.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002846:Chromatography, Affinity; D002855:Chromatography, Thin Layer; D010450:Endopeptidases / Q000378:metabolism*; D004789:Enzyme Activation; D005341:Fibrinolysin; D007531:Isoflurophate; D046911:Macromolecular Substances; D008970:Molecular Weight; D010958:Plasminogen / Q000378:metabolism*; D011485:Protein Binding; D011817:Rabbits; D012995:Solubility; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D000596:Amino Acids; D046911:Macromolecular Substances; D007531:Isoflurophate; D010958:Plasminogen; D010450:Endopeptidases; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator