Colocalization of gold-labeled LDL and fibrinogen on platelets: enhanced fibrinogen binding induced by LDL.
Low-density lipoproteins (LDL) specifically bind to the human platelet integrin-alpha IIb and -beta 3 (Koller et al., J. Biol. Chem. 264: 12412-12418, 1989). We show by electron microscopy (EM) that gold (Au)-labeled LDL bind to sites randomly distributed on the surface of platelets in suspension. Within a few minutes, mobile ligand-receptor complexes are translocated from the surface to the open canalicular system (OCS), which finally centralizes as a broad belt. Binding and translocation of Au-LDL are independent of stimulation of platelets by ADP and are completely reversible. Au-fibrinogen shows a strikingly similar, though agonist-dependent, redistribution behavior. Platelets are markedly activated by LDL. This activation is initiated by the binding of LDL to the plasma membrane receptor, and receptor internalization is probably not required for the activation but may instead be one of its consequences. Coincubation with Au-LDL and Au-fibrinogen results in more pronounced activation. The amount of OCS-localized ligands is significantly increased, most likely reflecting enhanced receptor recycling. The two ligands show a tendency to segregate in separate clusters, indicating differences in their postbinding pathways.
Volf
I
I
Institute of Medical Physiology, University of Vienna, Austria.
Koller
E
E
Bielek
E
E
Koller
F
F
D000244:Adenosine Diphosphate / Q000494:pharmacology; D001665:Binding Sites; D001667:Binding, Competitive; D001792:Blood Platelets / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D005340:Fibrinogen / Q000032:analysis / Q000378:metabolism*; D017979:Gold Colloid; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000032:analysis / Q000097:blood*; D015539:Platelet Activation; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism*; D011973:Receptors, LDL / Q000378:metabolism*; D013379:Substrate Specificity
D017979:Gold Colloid; D008077:Lipoproteins, LDL; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D011973:Receptors, LDL; D000244:Adenosine Diphosphate; D005340:Fibrinogen