Fibrinogen lysine residue A alpha 157 plays a crucial role in the fibrin-induced acceleration of plasminogen activation, catalyzed by tissue-type plasminogen activator.

M Voskuilen,G.H Veeneman,E.A Klasen,A Vermond,W Nieuwenhuizen,N.D Zegers,J.H Van Boom

doi:10.1016/s0021-9258(18)45518-4

Abstract

In previous studies, we have shown that the stretch 148-197 of the fibrinogen A alpha chain plays a crucial role in the acceleration of the tissue-type plasminogen activator (t-PA)-catalyzed plasminogen activation. In this study we have synthesized parts of A alpha 148-197 and analogues thereof. We found that the peptides with sequences identical with A alpha 148-161 and A alpha 149-161 of human fibrinogen accelerate the plasminogen activation by t-PA, whereas the corresponding peptides in which lysine residues A alpha 157 had been replaced by valine or arginine had no accelerating capacity. Furthermore, succinylation of the lysine residue(s) in the synthesized peptides A alpha 148-161 and A alpha 149-161 leads to loss of accelerating action. These findings show that lysine residue A alpha 157 is crucial for the accelerating action of fibrin on the t-PA-catalyzed plasminogen activation.

Highlights

Laboratory TNO, Rijswijk, The Netherlands from large scale melanoma cell culture according to Rijken et al [13], as modified by Kluft et al [14]
161 of human fibrinogen accelerate the plasminogen cording to the standard protocol using tert-butyloxycarbonyl-amino activation by t-PA, whereas the corresponding pep- acids with the following side-chain protection: Lys-2-chlorocarbotides in which lysine residues Aa157 had been replaced benzoxy, 4-methoxybenzenesulfonyl, Glu-benzyl,Asp-benzyl, Serby valine or arginine hadno accelerating capacity
Succinylation of the lysine residue(s) in the synthesized peptides Aa148-161 and Aa149-161 leads to loss of accelerating action. These findings show that lysine residue Aa157 is crucial for the accelerating action of fibrin on the t

Summary

Willem NieuwenhuizenSII

Laboratory, State University of Leiden, llMedica Biological Tissue-typePlasminogen Activator-Two-chain t-PA was purified. 161 of human fibrinogen accelerate the plasminogen cording to the standard protocol using tert-butyloxycarbonyl-amino activation by t-PA, whereas the corresponding pep- acids with the following side-chain protection: Lys-2-chlorocarbotides in which lysine residues Aa157 had been replaced benzoxy, 4-methoxybenzenesulfonyl, Glu-benzyl,Asp-benzyl, Serby valine or arginine hadno accelerating capacity. Succinylation of the lysine residue(s) in the synthesized peptides Aa148-161 and Aa149-161 leads to loss of accelerating action. These findings show that lysine residue Aa157 is crucial for the accelerating action of fibrin on the t-. When the addition of succinic anhydride was completed, solid hydroxylamine (Fluka) was added to a final concentration of 1 M,and acid; t-PA,tissue-type plasminogen activator; EGTA, [ethylene- the pHwas adjusted to 10with 2 N NaOH.

RESULTS

DISCUSSION

KRLEVDIDIVIRS RLEVDIDIVIRS

Stimulation factor