title: Localization of protein kinase A and vitronectin in resting platelets and their translocation onto fibrin fibers during clot formation.
abstract: Physiological stimulation of platelets with thrombin brings about the release of protein kinase A (PKA) into the plasma. In human blood, this kinase singles out and phosphorylates vitronectin (Vn), a multifunctional regulatory protein, which was proposed to play an important role in the control of fibrinolysis. Here we present immuno-cytochemical evidence to show: (i) that intact platelets possess on their surface an ecto-PKA which can preferentially phosphorylate Vn; (ii) that in the resting platelet, both the catalytic and the regulatory subunits of PKA are present on the platelet surface, in the surface-connected canalicular system, and within the alpha-granules of the platelets; (iii) that the process initiated upon platelet activation, which leads to the formation of fibrin fibers and consequently forms the fibrin net, is accompanied by a translocation of PKA, of Vn, and of PAI-1 onto the fibrin fibers. We propose that the localization and the translocation of these proteins in the fibrin net, together with our finding that PKA phosphorylation of Vn reduces its grip of PAI-1, can unleash PAI-1 in its free form. The free PAI-1 can then assume its latent (non inhibitory) conformation, allow plasminogen activators to trigger the formation of active plasmin, and to initiate fibrinolysis.
authors.affiliation: Department of Medical Biology, Saarland University, Homburg-Saar/Germany.
authors.forename: E
authors.initials: E
authors.lastname: Morgenstern
authors.forename: U
authors.initials: U
authors.lastname: Gnad
authors.forename: K T
authors.initials: KT
authors.lastname: Preissner
authors.forename: R
authors.initials: R
authors.lastname: Dierichs
authors.forename: A
authors.initials: A
authors.lastname: Belleli
authors.forename: A
authors.initials: A
authors.lastname: Chestukhin
authors.forename: I
authors.initials: I
authors.lastname: Schvartz
authors.forename: S
authors.initials: S
authors.lastname: Shaltiel
mesh_terms: D001692:Biological Transport; D001792:Blood Platelets / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004789:Enzyme Activation; D005337:Fibrin / Q000378:metabolism*; D006801:Humans; D010766:Phosphorylation; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism; D013997:Time Factors; D019096:Vitronectin / Q000378:metabolism*
chemical_list: D017395:Plasminogen Activator Inhibitor 1; D019096:Vitronectin; D005337:Fibrin; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases