Inhibition of plasminogen activation by apo(a): role of carboxyl-terminal lysines and identification of inhibitory domains in apo(a).
Apo(a), the distinguishing protein component of lipoprotein(a) [Lp(a)], exhibits sequence similarity to plasminogen and can inhibit binding of plasminogen to cell surfaces. Plasmin generated on the surface of vascular cells plays a role in cell migration and proliferation, two of the fibroproliferative inflammatory events that underlie atherosclerosis. The ability of apo(a) to inhibit pericellular plasminogen activation on vascular cells was therefore evaluated. Two isoforms of apo(a), 12K and 17K, were found to significantly decrease tissue-type plasminogen activator-mediated plasminogen activation on human umbilical vein endothelial cells (HUVECs) and THP-1 monocytes and macrophages. Lp(a) purified from human plasma decreased plasminogen activation on THP-1 monocytes and HUVECs but not on THP-1 macrophages. Removal of kringle V or the strong lysine binding site in kringle IV10 completely abolished the inhibitory effect of apo(a). Treatment with carboxypeptidase B to assess the roles of carboxyl-terminal lysines in cellular receptors leads in most cases to decreases in plasminogen activation as well as plasminogen and apo(a) binding; however, inhibition of plasminogen activation by apo(a) was unaffected. Our findings directly demonstrate that apo(a) inhibits pericellular plasminogen activation in all three cell types, although binding of apo(a) to cell-surface receptors containing carboxyl-terminal lysines does not appear to play a major role in the inhibition mechanism.
Department of Chemistry & Biochemistry, University of Windsor, Windsor, ON, Canada; and.
Rocco
R
Romagnuolo
Santica M
SM
Marcovina
Michael B
MB
Boffa
Marlys L
ML
Koschinsky
D001054:Apolipoproteins A / Q000737:chemistry / Q000502:physiology*; D002478:Cells, Cultured; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008239:Lysine / Q000502:physiology; D008264:Macrophages / Q000378:metabolism; D010958:Plasminogen / Q000502:physiology*; D010960:Plasminogen Activators / Q000737:chemistry / Q000494:pharmacology; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs
D001054:Apolipoproteins A; D010958:Plasminogen; D010960:Plasminogen Activators; D008239:Lysine
apolipoprotein(a); atherosclerosis; carboxypeptidase B; fibrinolysis; lipoprotein(a); thrombin-activatable fibrinolysis inhibitor; tissue-type plasminogen activator