Binding of plasmin (ogen) to rat C6 glioma cells is saturable and kringle-domain dependent. This interaction was studied as a model of plasimin(ogen) receptor interactions in nucleated mammalian cells. Apparent l25 I-plasmin dissociation from OS cell binding sites was slow; however, the dissociation rate was increased when the solution contained diisopropyl phosphoryl-plasmin (0.3 juM), fibrinogen (0.16 or 0.8 mg/ml), 1.08 mM D-Val-L-Leu-L-Lys-p-nitroanilide-HCl (S-2251), or e-amino-n-caproic acid (EACA, 5.0 mM). EACA promoted the most rapid dissociation of plasmin. C6 cell-associated plasmin and plasmin in solution demonstrated similar amidase activity. Only specifically bound plasmin (75% of total binding) was active against S-2251. Plasmin that was initially bound to C6 cells digested fibrinogen in a time-and plasmin concentration-dependent manner. o^-Antiplasmin (<* 2 AP, 0.1 fiM) completely inhibited fibrinogenolysis by plasmin that was initially C6 • or human umbilical vein endothelial-cell associated. Since a 2 AP reacts selectively with plasmin in solution (minimally with plasmin bound to cells), fibrinogen digestion by cell-associated plasmin probably occurred only after the plasmin dissociated into solution. Crosslinked fibrin clots were formed in uniform layers over C6 cells. If the cells were incubated with plasmin before addition of fibrinogen and thrombin, the clots were rapidly lysed.