Structural studies of fibrinolysis by electron microscopy.
Fibrin is degraded by the fibrinolytic system in which a plasminogen activator converts plasminogen to plasmin, a serine protease that cleaves specific bonds in fibrin leading to solubilization. To elucidate further the biophysical processes involved in conversion of insoluble fibers to soluble fragments, fibrin was treated with either plasmin or the combination of plasminogen and plasminogen activator, and morphologic changes were observed using scanning electron microscopy. These changes were correlated with biochemical analysis and with characterization of released, soluble fragments by transmission electron microscopy. Initial changes in the fibrin matrix included creation of many free fiber ends and gaps in the continuity of fibers. With more extensive digestion, free fiber segments associated laterally, resulting in formation of thick fiber bundles. Supernatants of digesting clots, containing soluble derivatives, were negatively contrasted and examined by transmission electron microscopy. Large, complex fragments containing portions of multiple fibers were observed, as were pieces of individual fibers and smaller fragments previously identified. Some large fragments had sharply defined ends, indicating that they had been cleaved perpendicularly to the fiber direction. Other fibers showed splayed ends or a lacy meshwork of surrounding protofibrils. Longer times generated more small fragments whose molecular composition could be inferred from their appearance. These results indicate that fibrinolytic degradation results in larger pieces than previously identified and that plasmin digestion proceeds locally by transverse cutting across fibers rather than by progressive cleavage uniformly around the fiber.
Veklich
Y
Y
Department of Cell and Developmental Biology, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.
Francis
C W
CW
White
J
J
Weisel
J W
JW
D004591:Electrophoresis, Polyacrylamide Gel; D005337:Fibrin / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D005341:Fibrinolysin / Q000494:pharmacology; D005342:Fibrinolysis / Q000502:physiology*; D005343:Fibrinolytic Agents / Q000494:pharmacology; D006801:Humans; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D010958:Plasminogen / Q000502:physiology; D013997:Time Factors; D010959:Tissue Plasminogen Activator / Q000494:pharmacology
D005343:Fibrinolytic Agents; D005337:Fibrin; D010958:Plasminogen; D010959:Tissue Plasminogen Activator; D005341:Fibrinolysin