Endogenous plasmin converts Glu-plasminogen to Lys-plasminogen on the monocytoid cell surface.
Recently, we showed that localization of Glu-plasminogen on cell surfaces enhances its conversion to Lys-plasminogen by exogenous plasmin. This leads to stimulation of plasminogen activation because Lys-plasminogen is the preferred substrate on cell surfaces. Here, we show that Glu-plasminogen was converted to Lys-plasminogen on monocytoid cells in the absence of exogenous plasmin. Culture of cells under serum-free conditions did not affect this conversion, suggesting that the enzymatic activity was cell-derived. Therefore, we tested whether endogenous monocytoid plasminogen could provide a source of plasmin to convert cell-associated Glu-plasminogen to Lys-plasminogen because plasmin is the only enzyme known to effect this reaction. We used a recombinant human plasminogen mutant, [D(646)E]Pg, which can be cleaved by plasminogen activators, but cannot catalyze the generation of Lys-plasminogen. Upon incubation with either THP-1 or U937 monocytoid cells, 35 and 38%, respectively, of the cell-bound ligand was converted to Lys-[D(646)E]Pg. Trasylol, alpha2-antiplasmin, and an anticatalytic antiplasminogen monoclonal antibody decreased Lys-[D(646)E]Pg formation to < 5% on monocytoid cells, consistent with a plasmin-dependent mechanism. Plasminogen was detected in these cells by Northern blotting and RT-PCR. Our results suggest that plasmin converts cell-bound Glu-plasminogen to Lys-plasminogen and that this enzyme is produced by activation of monocytoid plasminogen by endogenous monocytoid plasminogen activators to enhance plasminogen activation on the monocytoid cell surface.
Zhang
L
L
The Scripps Research Institute, CVN-26, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Gong
Y
Y
Grella
D K
DK
Castellino
F J
FJ
Miles
L A
LA
D002460:Cell Line; D004789:Enzyme Activation; D005341:Fibrinolysin / Q000378:metabolism*; D006801:Humans; D009000:Monocytes / Q000378:metabolism*; D009154:Mutation; D010446:Peptide Fragments / Q000096:biosynthesis; D010958:Plasminogen / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D010960:Plasminogen Activators / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D010446:Peptide Fragments; D012333:RNA, Messenger; D011994:Recombinant Proteins; C018998:lysyl-plasminogen; D010958:Plasminogen; D010960:Plasminogen Activators; D005341:Fibrinolysin