title: MMP-2 regulates human platelet activation by interacting with integrin alphaIIbbeta3.
abstract: BACKGROUND
  Human platelets contain matrix metalloproteinases (MMPs) that are secreted during platelet activation. Platelet MMPs have been implicated in the regulation of cellular activation and aggregation. Although the proaggregatory effect of MMP-2 has been demonstrated, the functional mechanism is not clearly understood.
  
  
  OBJECTIVE
  This work was carried out in order to elucidate the biochemical mechanism of MMP-2-associated platelet activation and aggregation.
  
  
  METHODS
  MMP-2 binding to the platelet surface was analyzed by flow cytometry. The cell surface target of MMP-2 was identified in thrombin receptor-activating peptide-stimulated platelets by immunoprecipitation, Western blotting and fluorescence microscopy. A recombinant hemopexin-like domain was used to characterize the nature of MMP-2 binding to the platelet surface. The functional significance of MMP-2 in platelet activation was investigated by quantitative measurements of the activation markers P-selectin (CD62P) and active alpha(IIb)beta(3). The role of MMP-2 in platelet aggregation was analyzed with an aggregometer.
  
  
  RESULTS
  ProMMP-2 binds to integrin alpha(IIb)beta(3) in stimulated platelets in which proMMP-2 is converted into MMP-2. Fibrinogen was able to replace the alpha(IIb)beta(3)-bound MMP-2. The molecular interaction of MMP-2 and integrin alpha(IIb)beta(3) was abrogated by the recombinant human hemopexin-like domain of MMP-2, leading to reduced cell surface expression of activation markers CD62P and active alpha(IIb)beta(3), and resulting in suppressed platelet aggregation.
  
  
  CONCLUSIONS
  This work clearly demonstrates that platelet activation and aggregation is regulated by MMP-2 that specifically interacts with integrin alpha(IIb)beta(3). The C-terminal hemopexin-like domain of MMP-2 is an essential element for binding to alpha(IIb)beta(3).
authors: National Research Laboratory, Department of Biochemistry, College of Science, Yonsei University, Seoul, Korea. dskim@yonsei.ac.kr
authors: W-S
authors: WS
authors: Choi
authors: O-H
authors: OH
authors: Jeon
authors: H-H
authors: HH
authors: Kim
authors: D-S
authors: DS
authors: Kim
mesh_terms: D020816:Amino Acid Motifs; D001792:Blood Platelets / Q000201:enzymology*; D020134:Catalytic Domain; D002462:Cell Membrane / Q000378:metabolism; D003094:Collagen / Q000737:chemistry; D005340:Fibrinogen / Q000737:chemistry; D015971:Gene Expression Regulation, Enzymologic; D006466:Hemopexin / Q000737:chemistry; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D008954:Models, Biological; D010446:Peptide Fragments / Q000737:chemistry; D015539:Platelet Activation; D010974:Platelet Aggregation; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism*
chemical_list: D010446:Peptide Fragments; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; C086309:thrombin receptor peptide SFLLRNP; D005340:Fibrinogen; D003094:Collagen; D006466:Hemopexin; D020778:Matrix Metalloproteinase 2