A comparison of bovine serum albumins, modified with a variety of carboxyl group agents, as stimulators of tissue-type plasminogen activator-catalyzed activation of plasminogen.
Bovine serum albumins (BSA), modified with a variety of carboxyl group agents, stimulated the tissue-type plasminogen activator (t-PA)-catalyzed activation of human plasminogen. Modification with taurine (tau) and putrescine (put) provided the best stimulants. The tauBSA and putBSA were effective at a concentration of 5 micrograms/ml and enhanced the Lys-plasminogen activation by two-chain t-PA in a dose-dependent manner to a maximum of 44- to 46-fold at 200 micrograms/ml. The Km values for the activation of Glu-plasminogen by t-PA in the presence of tauBSA and putBSA (100 micrograms/ml) were 1.7 and 1.8 microM, while the kcat values were 0.059 and 0.062 s-1, respectively. T-PA was bound to both tauBSA and putBSA, which were immobilized on agarose beads, with KD values of 163 and 138 nM, respectively. The two modified BSAs were good substrates for plasmin and were hydrolyzed by the enzyme to small peptides. All of these modified BSA-related actions were inhibited by lysine analogs (e.g. tranexamic acid) which were adjusted to the concentrations required for the inhibition of the plasminogen (Kringle 1 domain) binding to fibrin. On the other hand, acetylation or succinylation of the amino groups of BSA was not effective, while alkylation of the thiol groups of this protein resulted in a moderate stimulation of the plasmin generation. The present results show that t-PA and plasminogen form complexes with certain charge-modified BSAs via their lysine-binding sites. The different stimulation potency of modified BSAs may provide a model for in vivo counterparts of fibrin.
Soeda
S
S
Shimeno
H
H
Nagamatsu
A
A
D000818:Animals; D001711:Biotransformation / Q000187:drug effects; D002417:Cattle; D010958:Plasminogen / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000031:analogs & derivatives / Q000494:pharmacology*; D010959:Tissue Plasminogen Activator / Q000494:pharmacology*
D012710:Serum Albumin, Bovine; D010958:Plasminogen; D010959:Tissue Plasminogen Activator