Autoradiographic and immunoblotting analyses of the activation pathways of plasminogen by urokinase and tissue plasminogen activator in the plasma or clotted plasma

Takada, Akikazu; Yoshida, Masaya; Takada, Yumiko

doi:10.1016/0049-3848(88)90213-7

Cited by 4 publications

(2 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Since Glu-plasminogen is sensitive to plasmin and the activation rates of Lys-plasminogen are higher than those of the Glu-form, it has been proposed [3][4][5][6] that activation of plasminogen yields Lys-plasmin and requires the formation of Lysplasminogen as an intermediate. In most of the cases, however, a homogeneous purified system and urokinase (EC 3.4.21.31), an activator having no affinity for fibrin, have been used.…”

Section: Introductionmentioning

confidence: 99%

“…With the use of various types of fibrin mimics it has been recently proposed that the initial generation of Glu-plasmin by t-PA may proceed to the formation of Lys-plasmin by auto-catalysis [6,14] or through the formation of Lys-plasminogen as an intermediate [4,5,15]. Nevertheless, Holvoet et al [16] have recently suggested, on the basis of the absence of Lys-derived forms of plasminogen in the circulation, that activation of native Glu-plasminogen in vivo appears to occur by direct cleavage of the Arg-560-Val-561 bond in Glu-plasminogen and not via formation of Lys-plasminogen.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The mechanism of activation of plasminogen at the fibrin surface by tissue-type plasminogen activator in a plasma milieu in vitro. Role of α2-antiplasmin

Rouy

Anglès-Cano

1990

Biochemical Journal

View full text Add to dashboard Cite

The mechanism of activation of human Glu-plasminogen by fibrin-bound tissue-type plasminogen activator (t-PA) in a plasma environment or in a reconstituted system was characterized. A heterogeneous system was used, allowing the setting of experimental conditions as close as possible to the physiological fibrin/plasma interphase, and permitting the separate analysis of the products present in each of the phases as a function of time. The generation of plasmin was monitored both by spectrophotometric analysis and by radioisotopic analysis with a plasmin-selective chromogenic substrate and radiolabelled Glu-plasminogen respectively. Plasmin(ogen)-derived products were identified by SDS/PAGE followed by autoradiography and/or immunoblotting. When the activation was performed in a plasma environment, the products identified on the fibrin surface were Glu-plasmin (90%) and Glu-plasminogen (10%), whereas in the soluble phase only complexes between Glu-plasmin and its fast-acting inhibitor were detected. Identical results were obtained with a reconstituted system comprising solid-phase fibrin, t-PA, Glu-plasminogen and and alpha 2-antiplasmin. In contrast, when alpha 2-antiplasmin was omitted from the solution, Lys-plasmin was progressively generated on to the fibrin surface (30%) and released to the soluble phase. In the presence of alpha 2-antiplasmin or in plasma, the amount of active plasmin generated on the fibrin surface was lower than in the absence of the inhibitor: in a representative experiment the initial velocity of plasmin generation was 2.8 x 10(-3), 2.0 x 10(-3) and 1.8 x 10(-3) (delta A405/min) for 200 nM-plasminogen, 200 nM-plasminogen plus 100 nM-alpha 2-antiplasmin and native plasma respectively. Our results indicate that in plasma or in a reconstituted purified system containing plasminogen and alpha 2-antiplasmin at a ratio similar to that found in plasma (1) the activation pathway of native Glu-plasminogen proceeds directly to the formation of Glu-plasmin, (2) Lys-plasminogen is not an intermediate of the reaction and therefore (3) Lys-plasmin is not the final active product. However, in the absence of the inhibitor, Lys-plasmin and probably Lys-plasminogen, which is more readily activated to plasmin than is Glu-plasminogen, are generated as well.

show abstract

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The mechanism of activation of plasminogen at the fibrin surface by tissue-type plasminogen activator in a plasma milieu in vitro. Role of α2-antiplasmin

Rouy

Anglès-Cano

1990

Biochemical Journal

View full text Add to dashboard Cite

show abstract

Topically applied antithrombotic agents offer a new therapeutic approach to the prevention of microvascular thrombosis

Cooley

Gould

1991

Microsurgery

View full text Add to dashboard Cite

In the quest to develop optimal antithrombotic therapies for reconstructive microsurgery, with concomitant minimization of patient risk for generalized hemorrhage, surgeons have been turning to localized intraarterial delivery of various agents. An extension of this direction is to design agents that bind specifically to the site of thrombogenesis and effectively inhibit or prevent the buildup of thrombotic components. Progress in this direction must make use of the latest developments in the molecular understanding of coagulation, platelet adhesion/aggregation, and fibrinolytic processes. This article reviews pertinent developments in the biochemical understanding of thrombosis and discusses current avenues of investigation in the development of topically applied agents that help prevent microvascular thrombotic occlusion.

show abstract

The effects of polysaccharides on plasminogen activation by single chain- and two chain-tissue plasminogen activator

Kawamura

Watanabe

Urano

et al. 1991

Thrombosis Research

View full text Add to dashboard Cite

Autoradiographic and immunoblotting analyses of the activation pathways of plasminogen by urokinase and tissue plasminogen activator in the plasma or clotted plasma

Cited by 4 publications

References 16 publications

The mechanism of activation of plasminogen at the fibrin surface by tissue-type plasminogen activator in a plasma milieu in vitro. Role of α2-antiplasmin

The mechanism of activation of plasminogen at the fibrin surface by tissue-type plasminogen activator in a plasma milieu in vitro. Role of α2-antiplasmin

Topically applied antithrombotic agents offer a new therapeutic approach to the prevention of microvascular thrombosis

The effects of polysaccharides on plasminogen activation by single chain- and two chain-tissue plasminogen activator

Contact Info

Product

Resources

About