Subcellular distribution of fibrinogen and factor XIII in human blood platelets

Łopaciuk, S; Lovette, K.M; McDonagh, Jan; Chuang, Hanson Y. K.; McDonagh, R. P.

doi:10.1016/0049-3848(76)90223-1

Cited by 84 publications

(46 citation statements)

References 20 publications

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“…FXIII has previously been detected in platelet releasates, 54,55 but these observations are not consistent with our data or other reports. 23,24,37,46 FXIII-A levels are normal in platelets derived from patients with gray-platelet syndrome 25 and plasma FXIII levels are unchanged in thrombocytopenic mice.38 Together these data indicate a very minor contribution of a-granule FXIII-A 2 B 2 to the overall TG activity derived from platelet preparations.Platelet activation is essential for FXIII-A-mediated stabilization of FXIII-depleted thrombi. FXIII-A activity assays on platelets preactivated with TRAP-6/collagen are carried out in the absence of exogenous thrombin, suggesting that FXIII-A is nonproteolytically activated by the high Ca 21 levels associated with platelet stimulation.…”

supporting

confidence: 48%

“…Platelet FXIII-A, also known as cellular FXIII, 30 is largely stored in the cytoplasm 23,24 and, in contrast to FXIII-A 2 B 2 , can be nonproteolytically activated by Ca 21 alone. 31 Platelet-rich clots in vivo are much more resistant to lysis than whole blood clots.…”

mentioning

confidence: 99%

“…These inhibitors include a 2 -antiplasmin (a 2 AP), 17 thrombin activatable fibrinolysis inhibitor, 18 and plasminogen activator inhibitor-2. 19 a 2 AP is a serpin inhibitor that forms an irreversible complex with plasmin 20 and interferes with binding of plasminogen to fibrin.21 FXIIIa cross-links Gln-2 of a 2 AP to Lys-303 of the Aa chain of fibrin(ogen).22 The crucial role of crosslinked a 2 AP in stabilizing fibrin is demonstrated by the rapid lysis of clots formed in the absence of a 2 AP, 1,3,17 or when a 2 AP activity is neutralized.1 FXIII-A is present in platelets [22][23][24][25][26][27] in large quantities; a single platelet contains 60 6 10 fg, 28 making local platelet FXIII-A concentration around 150-fold greater than that in plasma. …”

mentioning

confidence: 99%

“…1 FXIII-A is present in platelets [22][23][24][25][26][27] in large quantities; a single platelet contains 60 6 10 fg, 28 making local platelet FXIII-A concentration around 150-fold greater than that in plasma.…”

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confidence: 99%

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Functional factor XIII-A is exposed on the stimulated platelet surface

Mitchell

Lionikiene

Fraser

et al. 2014

Blood

105

141

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Key Points• Factor XIII-A is exposed in protruding caps on the activated platelet surface.• Platelet FXIII-A exerts antifibrinolytic function by cross-linking a 2 AP to fibrin.Factor XIII (FXIII) stabilizes thrombi against fibrinolysis by cross-linking a 2-antiplasmin (a 2 AP) to fibrin. Cellular FXIII (FXIII-A) is abundant in platelets, but the extracellular functions of this pool are unclear because it is not released by classical secretion mechanisms. We examined the function of platelet FXIII-A using Chandler model thrombi formed from FXIIIdepleted plasma. Platelets stabilized FXIII-depleted thrombi in a transglutaminase-dependent manner. FXIII-A activity on activated platelets was unstable and was rapidly lost over 1 hour. Inhibiting platelet activation abrogated the ability of platelets to stabilize thrombi. Incorporating a neutralizing antibody to a 2 AP into FXIII-depleted thrombi revealed that the stabilizing effect of platelet FXIII-A on lysis was a 2 AP dependent. Platelet FXIII-A activity and antigen were associated with the cytoplasm and membrane fraction of unstimulated platelets, and these fractions were functional in stabilizing FXIII-depleted thrombi against lysis. Fluorescence confocal microscopy and flow cytometry revealed exposure of FXIII-A on activated membranes, with maximal signal detected with thrombin and collagen stimulation. FXIII-A was evident in protruding caps on the surface of phosphatidylserine-positive platelets. Our data show a functional role for platelet FXIII-A through exposure on the activated platelet membrane where it exerts antifibrinolytic function by cross-linking a 2 AP to fibrin. (Blood. 2014;124(26):3982-3990) IntroductionFactor XIII (FXIII) plays an essential role in normal hemostasis where it contributes to the regulation of fibrinolysis, 1-3 the maintenance of pregnancy, 4 wound healing, and angiogenesis. 5 The function of FXIII in hemostasis is further emphasized in its deficiency, which results in hemorrhage 6,7 and slow wound healing. 4,7 FXIII circulates in plasma as a heterotetramer (FXIII-A 2 B 2 ) where the catalytic A (FXIII-A 2 ) subunits are almost exclusively in complex with the inhibitory carrier B (FXIII-B 2 ) subunits. 8 FXIII is activated by the combined action of thrombin and Ca 21 to form the active transglutaminase (TG) FXIIIa. 9,10 FXIIIa confers stability to the fibrin matrix by cross-linking fibrin, substantially altering its rheologic properties.11,12 TG catalyzes formation of covalent e-(g-glutamyl) lysyl bonds 13 in which the lysine e-amino group is cross-linked to the glutamine g-carboxymide group. 14The cross-linking of g-chains confers a degree of rigidity to the fibrin network, which is further stabilized by the cross-linking of a-chains 15 to generate high-molecular-weight polymers. 10,16 FXIIIa also cross-links inhibitors of fibrinolysis to fibrin, further increasing its insolubility and resistance to plasmin. These inhibitors include a 2 -antiplasmin (a 2 AP), 17 thrombin activatable fibrinolysis inhibitor, 18 and plasminogen ac...

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supporting

confidence: 48%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Functional factor XIII-A is exposed on the stimulated platelet surface

Mitchell

Lionikiene

Fraser

et al. 2014

Blood

105

141

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“…20 Although this factor XHIa is presumably of platelet origin, it is unlikely that the platelet has a mechanism to move this protein through the plasma membrane from its cytoplasmic location. 21 It is therefore probable that platelet-associated factor XIII either is of plasma origin or is derived from lysed platelets. The mechanism of the association between platelets and factor Xllla is unexplored.…”

Section: Discussionmentioning

confidence: 99%

Platelet-associated factor XIII as a marker of platelet activation in patients with peripheral vascular disease.

Devine

Andestad

Nugent

et al. 1993

Arterioscler Thromb

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In the past several years, monoclonal antibodies have been developed that distinguish between resting and activated platelets IN vitro. These antibodies recognize epitopes expressed on membrane proteins or soluble proteins, such as factor XIHa and thrombospondin, that bind only to activated platelets. We have used fluorescence flow cytometry to determine whether three such antibodies can detect platelet activation in patients with severe peripheral vascular disease (FVD). Using two activation-specific monoclonal antibodies and a polyclonal antiserum to factor XIII a-chain, we have examined the platelets from PVD patients, age-matched control subjects who were free of detectable PVD, and unmatched control subjects. Cells analyzed as platelets were identified by their light-scatter profile and their reactivity with monoclonal anti-glycoprotein Ib. The platelets of patients with PVD showed no increase in binding of activation-dependent 1B3 (directed against a 180-kD membrane protein) compared with age-matched control subjects (p=0.780). Similarly, there was no difference between PVD patients and control subjects in activation-dependent CD63 expression. Conversely, the binding of anti-factor XIII a-chain was significantly higher than in the control groups (/><0.001). These data suggest that the detection of soluble factors that bind to activated but not resting platelets may be of use in the detection of pathological in vivo platelet activation.

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Identification of fibrinogen derivatives in the triton‐insoluble residue of human blood platelets

et al. 1983

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Several proteins (eg. actin, myosin, and actin-binding protein) in the Triton-insoluble residue of thrombin-stimulated platelets are important in the formation of cytoskeletal structures. Electrophoretic analyses have shown that unidentified protein bands of 68,000, 55,000, and 48-50,000 daltons are also present in larger amounts after thrombin stimulation. Since these molecular weights correspond roughly to those of the alpha, beta, and gamma chains of fibrin, and since fibrinogen is found in platelet alpha-granules, these bands were compared to those obtained when purified fibrinogen was treated with thrombin, exposed to 1% Triton X-100-5 mM EGTA, and the resultant Triton-insoluble residue sedimented. Identification of the 68,000-, 55,000-, and 48-50,000-dalton bands as fibrinogen derivatives was confirmed by identifying them in comigration studies and in autoradiographs of Triton-insoluble residues of platelets that were electrophoretically transferred to nitrocellulose paper and treated with antifibrinogen antibody and 125I-protein A. Furthermore, if the platelet suspension was treated with thrombin in the presence of calcium ions, protein bands characteristic of the action of Factor XIII on fibrin were observed, active platelet Factor XIII apparently having been made available by lysis of platelets during preparations. Making use of the electrophoretic properties of tubulin recently described by Best et al [1981], comigration studies using hog brain tubulin indicated that tubulin is not present in significant amounts in the Triton-insoluble residue of platelets as previously suggested. The identification of these proteins as fibrinogen derivatives does not demonstrate a physiological interaction between fibrin and the platelet cytoskeleton, since fibrin is Triton-insoluble and can be pelleted even in the absence of platelet cytoskeletons.

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Subcellular distribution of fibrinogen and factor XIII in human blood platelets

Cited by 84 publications

References 20 publications

Functional factor XIII-A is exposed on the stimulated platelet surface

Functional factor XIII-A is exposed on the stimulated platelet surface

Platelet-associated factor XIII as a marker of platelet activation in patients with peripheral vascular disease.

Identification of fibrinogen derivatives in the triton‐insoluble residue of human blood platelets

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