Defensin modulates tissue-type plasminogen activator and plasminogen binding to fibrin and endothelial cells.
Defensins are naturally occurring antimicrobial peptides that may participate in host defense against microorganisms. We previously reported that the amino acid sequence of leukocyte defensins resembles the lysine-binding site in the kringles of plasminogen and that defensin inhibits fibrinolysis mediated by tissue-type plasminogen activator (tPA) and plasminogen. In the present paper we analyze the mechanisms of this inhibition. Defensin binds specifically to cultured human umbilical vein endothelial cells (HUVEC) (half-maximal binding = 3 microM) as well as to fibrin. At saturating concentrations (5-10 microM), defensin stimulates the maximum binding of plasminogen to HUVEC and to fibrin approximately 10-fold. However, defensin inhibits plasminogen binding to both surfaces at concentrations >10 microM. Defensin also inhibits tPA and plasminogen-mediated fibrinolysis in a dose-dependent manner at all concentrations tested. Fibrinolysis is almost totally inhibited by 6 microM defensin, a concentration that stimulates the binding of plasminogen to fibrin. Discordance between the enhancement of plasminogen binding and its activation cannot be explained by an inhibitory effect of defensin on tPA binding nor by inhibition of plasmin activity, each of which occur only at higher concentrations. Rather, these results suggest that plasminogen bound to fibrin in the presence of defensin is less susceptible to activation by tPA.
Higazi
A A
AA
Department of Pathology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Ganz
T
T
Kariko
K
K
Cines
D B
DB
D000890:Anti-Infective Agents / Q000494:pharmacology*; D001798:Blood Proteins / Q000494:pharmacology*; D023082:Defensins; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000378:metabolism*; D005337:Fibrin / Q000378:metabolism*; D006801:Humans; D007962:Leukocytes / Q000737:chemistry; D010958:Plasminogen / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D010959:Tissue Plasminogen Activator / Q000378:metabolism*
D000890:Anti-Infective Agents; D001798:Blood Proteins; D023082:Defensins; D005337:Fibrin; D010958:Plasminogen; D010959:Tissue Plasminogen Activator