Agglutination of isolated platelet membranes.
Platelet membrane glycoproteins play a central role in platelet aggregation and thus in primary hemostasis. To investigate mechanisms of platelet-platelet interaction in the absence of cellular activation events, we studied immunological and functional aspects of isolated platelet membranes. Platelet membranes contained significant amounts of the inducible fibrinogen receptor, glycoprotein (GP) IIb-IIIa, which exposes conformation-dependent LIBS1 and PMI-1 epitopes in response to fibrinogen-mimetic peptides GRGDSP and HHLG-GAKQAGDV. In the presence of soluble fibrinogen, membrane-coated latex beads showed Ca(2+)-dependent agglutination that could be partially inhibited by GRGDSP but not by the biologically inactive peptide GRGESP. Thrombospondin enhanced agglutination of membrane-coated beads, which could be inhibited by polyvalent anti-thrombospondin Fab fragments and anti-thrombospondin monoclonal antibody MA-II. Mg2+ inhibited both GPIIb-IIIa- and thrombospondin-mediated agglutination of membranes in a dose-dependent manner. The results of the present study indicate that isolated platelet membranes are a useful tool to study regulation of GPIIb-IIIa- and thrombospondin-mediated platelet-platelet interaction.
Gawaz
M
M
I Medizinische Klinik, Technische Universität München, Germany.
Ott
I
I
Reininger
A J
AJ
Heinzmann
U
U
Neumann
F J
FJ
D000371:Agglutination / Q000187:drug effects; D000911:Antibodies, Monoclonal; D001792:Blood Platelets / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D002413:Cations, Divalent / Q000494:pharmacology; D002462:Cell Membrane / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D015151:Immunoblotting; D008565:Membrane Proteins / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000502:physiology
D000911:Antibodies, Monoclonal; D002413:Cations, Divalent; D008565:Membrane Proteins; D019039:Platelet Glycoprotein GPIIb-IIIa Complex