Internalization of bound fibrinogen modulates platelet aggregation.
In agonist-stimulated platelets, the integrin alpha IIb beta 3 (glycoprotein IIb-IIIa) is converted from an inactive to an active fibrinogen receptor, thereby mediating platelet aggregation. With time after agonist addition, at least two events occur: fibrinogen becomes irreversibly bound to the platelet and, when stirring is delayed, platelets lose the ability to aggregate despite the presence of maximally bound fibrinogen. Because we previously identified an actively internalized pool of alpha IIb, beta 3 in platelets, we explored the possibility that both of these events might result from the internalization of fibrinogen bound to active alpha IIb beta 3. Under conditions of irreversible fibrinogen binding, fluorescence microscopy showed that biotinylated fibrinogen is rapidly internalized by activated platelets to a surface-inaccessible, intracellular pool. Flow cytometric analysis showed that the observed loss in accessibility to extracellular probes immediately precedes a loss in ability to the platelets to aggregate. Moreover, prevention of irreversible fibrinogen binding results in a prevention of internalization and a retention of aggregation capacity. Thus, the internalization of fibrinogen from the activated platelet surface appears to contribute not only to the irreversible phase of fibrinogen binding, but also to the downregulation of platelet adhesiveness. Fibrinogen internalization is therefore likely to represent a fundamental regulatory mechanism that modulates platelet function.
Wencel-Drake
J D
JD
Department of Medical Laboratory Sciences, University of Illinois at Chicago 60612, USA.
Boudignon-Proudhon
C
C
Dieter
M G
MG
Criss
A B
AB
Parise
L V
LV
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000595:Amino Acid Sequence; D002451:Cell Compartmentation; D004705:Endocytosis; D005340:Fibrinogen / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000494:pharmacology; D010974:Platelet Aggregation / Q000187:drug effects / Q000502:physiology*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism*; D011485:Protein Binding; D015032:Zinc / Q000494:pharmacology
D010446:Peptide Fragments; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; C072032:thrombin receptor peptide (42-55); D000244:Adenosine Diphosphate; D005340:Fibrinogen; D015032:Zinc