Characterization of plasminogen as an adhesive ligand for integrins alphaMbeta2 (Mac-1) and alpha5beta1 (VLA-5).
Plasminogen (Pg) has been implicated in many biologic processes involving extracellular proteolysis. We investigated whether Pg, by virtue of its capacity to be deposited within the extracellular matrix, can serve as a ligand for cell surface integrins. We report here that Pg supports cell adhesion by engaging integrins alphaMbeta2 and alpha5beta1. The immobilized Glu-Pg, but not its derivatives with the N-terminal peptide lacking, plasmin and Lys-Pg, supported efficient adhesion that was abolished by anti-alphaMbeta2 and anti-alpha5beta1 integrin-specific monoclonal antibodies (mAbs). In addition, lysine binding sites of Glu-Pg contributed to cell adhesion inasmuch as tranexamic acid and epsilon-aminocaproic acid inhibited cell adhesion. The involvement of alphaMbeta2 and alpha5)beta1 in adhesion to Glu-Pg was demonstrable with blood neutrophils, U937 monocytoid cells, and genetically engineered alphaMbeta2-transfected human embryonic kidney (HEK) 293 cells. In alphaMbeta2, the alphaMI-domain is the binding site for Glu-Pg because the "I-less" form of alphaMbeta2 did not support cell adhesion and the recombinant alphaMI-domain bound Glu-Pg directly. In comparison with cell adhesion, the binding of soluble Glu-Pg to cells and the concomitant generation of plasmin activity was inhibited by anti-alpha5beta1 but not by anti-alphaMbeta2. These findings identify Glu-Pg as an adhesive ligand for integrins alphaMbeta2 and alpha5beta1 and suggest that alpha5beta1 may participate in the binding of soluble Glu-Pg and assist in its activation.
Lishko
Valeryi K
VK
Joseph J. Jacobs Center for Thrombosis and Vascular Biology, Department of Molecular Cardiology, Lerner Research Institute, Cleveland, OH 44195, USA.
Novokhatny
Valery V
VV
Yakubenko
Valentin P
VP
Skomorovska-Prokvolit
Helen V
HV
Ugarova
Tatiana P
TP
D000595:Amino Acid Sequence; D002448:Cell Adhesion / Q000502:physiology*; D002460:Cell Line; D005341:Fibrinolysin / Q000378:metabolism; D005347:Fibroblasts / Q000502:physiology; D006801:Humans; D039081:Integrin alpha5beta1 / Q000502:physiology*; D007668:Kidney; D008024:Ligands; D016177:Macrophage-1 Antigen / Q000502:physiology*; D008969:Molecular Sequence Data; D010446:Peptide Fragments; D010958:Plasminogen / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D020349:Surface Plasmon Resonance; D014162:Transfection; D020298:U937 Cells
D039081:Integrin alpha5beta1; D008024:Ligands; D016177:Macrophage-1 Antigen; D010446:Peptide Fragments; D010958:Plasminogen; D005341:Fibrinolysin