Plasmic degradation of human fibrinogen I. Structural characterization of degradation products

Furlan, Miha; Beck, Eugene A.

doi:10.1016/0005-2795(72)90044-x

Cited by 82 publications

(28 citation statements)

References 12 publications

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“…sequence of degradation is similar to that observed in plasmin-catalyzed digests, in that Aa-chain fragmentation precedes that of BP-chain (12,(14)(15)(16)(17)(18) Fibrinogen Degradation by a2-Macroglobulin globulin fromi urokinase-treated plasilma (500 U urokinase/ml plasmiia). Less than 0.1% of this plasimin was expresse(c as fibinogenolvtic activity.…”

Section: Resultssupporting

confidence: 64%

“…Furthermore, the suggestion that the reduction in molecular size wvas the consequence of cleavage of peptide material from the COOH-terminal region of the core imiolecule (10,11) has been confirimied (12) and extended by the demonstration that uinder physiologic circumstances such attack appears to be conifined to the Aa-chains (12,14). Sinillarities between the plasmna catabolites and early fibrinogen derivatives resulting from plasmin-catalyzed hydrolysis (14)(15)(16)(17)(18) support the notion that it is plasminii or a plasmin-like enzyme that mediates this in vivo cataholismll.…”

Section: Introductionmentioning

confidence: 99%

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Degradation of Human Fibrinogen by Plasma α2-Macroglobulin-Enzyme Complexes

Harpel¹,

Mosesson²

1973

J. Clin. Invest.

159

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A B S T R A C T This study demonstrates that human plasma a2-macroglobulin preparations possess an enzymic activity that degrades fibrinogen, resulting in the formation of products whose structure resembles that of circulating fibrinogen catabolites. The sequence of degradation is similar to that observed in plasmincatalyzed digests, in that Aa-chain fragmentation precedes that of B,8-chain. The addition of plasminogen activators to plasma induced an increase in the N-atosyl-L-arginine methyl ester HCl esterase and fibrinogenolytic activity associated with a2-macroglobulin purified from this plasma, indicating that the enzymic activity of the complex was preserved and could be increased in the presence of other plasma enzyme inhibitors. Immunochemical studies demonstrated that an a2-macroglobulin-plasmin complex had formed in urokinase-treated plasma. This a2-macroglobulin preparation manifested an esterolytic profile like that of a complex prepared from plasmin and purified a2-macroglobulin. After complex formation with a2-macroglobulin in plasma, plasmin retained less than 0.1% of its fibrinogenolytic activity. That plasmin expressed its activity while bound to a2-macroglobulin was suggested by immunoprecipitation of this activity with aa-macroglobulin antibody and by the demonstration that pancreatic trypsin inhibitor did not effectively inhibit its fibrinogenolytic or esterolytic activity. These results raise the possibility that, in addition to its activity as a major plasma proteolytic enzyme inhibitor, a2-macroglobulin may modThis paper was presented in part at the 15th Annual Meeting,

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Section: Resultssupporting

confidence: 64%

Section: Introductionmentioning

confidence: 99%

Degradation of Human Fibrinogen by Plasma α2-Macroglobulin-Enzyme Complexes

Harpel¹,

Mosesson²

1973

J. Clin. Invest.

159

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“…Fig. 5 shows diagrammatically the covalently bound derivatives which would result from cleavages at known sites of plasmin action on fibrin (33,(38)(39)(40)(41)(42)(43)(44) 4 880,000 19.9 850,000 * Complexes named according to the elution peaks from Sephacrvl S-300 column (Fig. 4) each of which corresponds to bands 1-4 noted in Figs.…”

Section: Discussionmentioning

confidence: 99%

Plasmic Degradation of Crosslinked Fibrin

Francis¹,

Marder²,

Barlow³

1980

J. Clin. Invest.

123

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A B S T R A C T Crosslinked fibrin was digested by plasmin, and three soluble complexes larger than DD/E were purified and characterized. After gel filtration chromatography, the purified complexes were shown to have molecular weights of 465,000, 703,000, and 850,000, as determined by equilibrium sedimentation. Each of the complexes was dissociated into two or more fragments by SDS-polyacrylamide gel electrophoresis. The structure of these subunit fragments was deduced from determinations of their molecular weights and polypeptide chain composition and from known sites of plasmin cleavage of fibrin. Fragments larger than DD have been identified that contain intact yry crosslinks as well as fragments resulting from cleavages at or near this site. The former include DY (mol wt 247,000), YY (mol wt 285,000), DXD (mol wt 461,000), and YXD (mol wt 500,000); and the latter include fragments XD (mol wt 334,000) and XY (mol wt 391,000). A schematic model was developed to explain the structure of the large noncovalently bound complexes based on their molecular weight and observed component fragments. Our scheme supports the twostranded half-staggered overlap model as the basic unit of fibrin structure, in which each complex consists of fragments from two adjacent complementary antiparallel fibrin strands. The smallest derivative, complex 1, is the DD/E complex; complex 2 contains apposed DY and YD fragments, and complex 3 consists of fragments DXD and YY. Complex 4 is less wellcharacterized, but its intact structure is projected to consist of YXD and DXY fragments from adjacent fibrin strands. Each complex is heterogeneous in subunit composition, reflecting additional plasmin cleavages within and/or adjacent to its theoretical boundaries. Since most ofthe protein initially released into solution from degrading fibrin is as complexes larger than DD/E,

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“…Fractions of 5 ml were collected. When analyzed by nonreducing SDS-PAGE (24), the first emerging protein peak was found to comprise intact fibrinogen, X and Y fragments (31)(32)(33)(34), whereas the early portion of the second major peak showed a mixture of D and E fragments (32)(33)(34)(35). The last three fractions of this peak, however, contained D fragments alone.…”

Section: Methodsmentioning

confidence: 99%

Contact with the N Termini in the Central E Domain Enhances the Reactivities of the Distal D Domains of Fibrin to Factor XIIIa

Samokhin

Lóránd

1995

Journal of Biological Chemistry

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The reaction of Factor XIII a with fibrin is the last enzyme-catalyzed step on the coagulation cascade, leading to the formation of a normal blood clot. The finding that fibrin is preferred by the cross-linking enzyme about 10-fold over the circulating fibrinogen suggests the operation of a unique substrate-level control for the orderly functioning of the physiological process in the forward direction. An important task is to elucidate the molecular mechanism for the transmission of the signal generated by the thrombin-catalyzed cleavage in the central E domain of fibrin to the distant Factor XIII areactive glutamine residues. By focusing on the substrate sites present in ␥ chain remnants of D type domains of fibrinogen and by employing the approach of fragment complementation with the regulatory E domain, which represents the thrombin-modified portion of fibrin, we have now succeeded in reconstructing in solution the phenomenon of kinetic enhancement for the reaction with Factor XIII a .Two D type preparations (truncated fibrinogen, ϳ250 kDa and D, ϳ105 kDa) were obtained by digestion of human fibrinogen with endo Lys-C. Neither product could be cross-linked by Factor XIII a , but as shown by the incorporation of dansylcadaverine, both were acceptor substrates for the enzyme. The plasmin-derived D (ϳ105-kDa) product, however, could be cross-linked into DD dimers. In all cases, the admixture of E fragments exerted a remarkable boosting effect on the reactions with Factor XIII a . Even with native fibrinogen as substrate, cross-linking of ␥ chains was enhanced in the presence of E. Nondenaturing electrophoresis was used to demonstrate the complex forming potential of E fragments with fibrinogen, truncated fibrinogen, D, or D. The GPRP tetrapeptide mimic of the GPRV N-terminal sequence of the ␣ chains in the E fragments, abolished both complex formation and the kinetic boosting effect of E on the reactions of substrates with Factor XIII a . Thus, the N-terminal ␣ chain sequences seem to act as organizing templates for spatially orienting the D domains, probably during the protofibrillar assembly of the fibrin units, for favorable reaction with Factor XIII a .

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Plasmic degradation of human fibrinogen I. Structural characterization of degradation products

Cited by 82 publications

References 12 publications

Degradation of Human Fibrinogen by Plasma α2-Macroglobulin-Enzyme Complexes

Degradation of Human Fibrinogen by Plasma α2-Macroglobulin-Enzyme Complexes

Plasmic Degradation of Crosslinked Fibrin

Contact with the N Termini in the Central E Domain Enhances the Reactivities of the Distal D Domains of Fibrin to Factor XIIIa

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