scholarly journals Regulation of plasma factor XIII binding to fibrin in vitro

Blood ◽  
1985 ◽  
Vol 66 (5) ◽  
pp. 1028-1034 ◽  
Author(s):  
CS Greenberg ◽  
JV Dobson ◽  
CC Miraglia
Keyword(s):  
Factor Xiii ◽  
Specific Binding ◽  
Divalent Cations ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protamine Sulfate ◽  
Factor Xiiia ◽  
Plasma Sodium ◽  
Platelet Factor ◽  
Plasma Factor

Abstract The binding of plasma factor XIII to fibrinogen or fibrin that has been chemically or enzymatically induced to polymerize was studied. Factor XIII binding was assayed using a 3H-putrescine incorporation assay and an 125I-plasma factor XIII binding assay. More than 80% of the native and radiolabeled plasma factor XIII was bound to fibrin I formed by reptilase in EDTA, citrate, or heparin anticoagulated plasma. Plasma factor XIII and 125I-factor XIII was bound (89.6% to 92.5%) to fibrin II formed by thrombin in either citrate or EDTA anticoagulated plasma. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of 125I-plasma factor XIII bound to fibrin I or fibrin II formed by reptilase or thrombin in the presence of EDTA demonstrated the b2- subunit remained bound to the a-chains or thrombin-cleaved a-chains. In the presence of calcium chloride and thrombin, the b2-subunit dissociated and factor XIIIa was bound. Protamine sulfate caused fibrinogen polymerization in the absence of divalent cations and reduced both plasma factor XIII and immunologic fibrinogen levels. Fibrinogen polymerized by protamine sulfate bound plasma factor XIII and the a2-subunit of 125I-platelet factor XIII. Plasma factor XIII was also bound to sonicated non-cross-linked fibrin II in either normal plasma or afibrinogenemic plasma. Plasma levels of several coagulation proteins were unchanged after the addition of reptilase, protamine sulfate, or sonicated fibrin to plasma. These results demonstrate that a specific binding site for the a2-subunit of plasma factor XIII is present on polymerized fibrinogen, fibrin I, and fibrin II. Furthermore, the presence of divalent cations, thrombin-cleavage of plasma factor XIII, and release of fibrinopeptides A or B are not required for plasma factor XIII binding to polymerized fibrinogen and fibrin.

scholarly journals Regulation of plasma factor XIII binding to fibrin in vitro

Blood ◽  
1985 ◽  
Vol 66 (5) ◽  
pp. 1028-1034 ◽  
Author(s):  
CS Greenberg ◽  
JV Dobson ◽  
CC Miraglia
Keyword(s):  
Factor Xiii ◽  
Specific Binding ◽  
Divalent Cations ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protamine Sulfate ◽  
Factor Xiiia ◽  
Plasma Sodium ◽  
Platelet Factor ◽  
Plasma Factor

The binding of plasma factor XIII to fibrinogen or fibrin that has been chemically or enzymatically induced to polymerize was studied. Factor XIII binding was assayed using a 3H-putrescine incorporation assay and an 125I-plasma factor XIII binding assay. More than 80% of the native and radiolabeled plasma factor XIII was bound to fibrin I formed by reptilase in EDTA, citrate, or heparin anticoagulated plasma. Plasma factor XIII and 125I-factor XIII was bound (89.6% to 92.5%) to fibrin II formed by thrombin in either citrate or EDTA anticoagulated plasma. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of 125I-plasma factor XIII bound to fibrin I or fibrin II formed by reptilase or thrombin in the presence of EDTA demonstrated the b2- subunit remained bound to the a-chains or thrombin-cleaved a-chains. In the presence of calcium chloride and thrombin, the b2-subunit dissociated and factor XIIIa was bound. Protamine sulfate caused fibrinogen polymerization in the absence of divalent cations and reduced both plasma factor XIII and immunologic fibrinogen levels. Fibrinogen polymerized by protamine sulfate bound plasma factor XIII and the a2-subunit of 125I-platelet factor XIII. Plasma factor XIII was also bound to sonicated non-cross-linked fibrin II in either normal plasma or afibrinogenemic plasma. Plasma levels of several coagulation proteins were unchanged after the addition of reptilase, protamine sulfate, or sonicated fibrin to plasma. These results demonstrate that a specific binding site for the a2-subunit of plasma factor XIII is present on polymerized fibrinogen, fibrin I, and fibrin II. Furthermore, the presence of divalent cations, thrombin-cleavage of plasma factor XIII, and release of fibrinopeptides A or B are not required for plasma factor XIII binding to polymerized fibrinogen and fibrin.

scholarly journals Isolation of a fibrin-binding fragment from blood coagulation factor XIII capable of cross-linking fibrin(ogen)

1988 ◽  
Vol 256 (3) ◽  
pp. 1013-1019 ◽  
Author(s):  
C S Greenberg ◽  
J J Enghild ◽  
A Mary ◽  
J V Dobson ◽  
K E Achyuthan
Keyword(s):  
Active Site ◽  
Factor Xiii ◽  
Polyacrylamide Gel Electrophoresis ◽  
Limited Proteolysis ◽  
Coagulation Factor ◽  
Cysteine Residue ◽  
Factor Xiiia ◽  
Platelet Factor ◽  
Transglutaminase Activity ◽  
A Chain

Purified platelet Factor XIII was radioiodinated and then partially degraded by thrombin or trypsin, and a fibrin-binding fragment was identified by autoradiography and immunoblotting following separation by SDS/polyacrylamide-gel electrophoresis. Limited proteolysis of 125I-Factor XIII by thrombin or trypsin produced an 125I-51 kDa fragment and an unlabelled 19 kDa fragment. The 51 kDa fragment was purified by h.p.l.c. on a TSK-125 gel-filtration column. Partial amino acid sequence analysis of the 51 kDa fragment indicated that it was similar in sequence to the Gly38-Lys513 segment in placental Factor XIII a-chain. More than 70% of the 51 kDa fragment bound to fibrin, whereas the 19 kDa fragment did not bind. The active site was localized to the 51 kDa fragment since this fragment expressed transglutaminase activity, cross-linked fibrin and fibrinogen and incorporated iodo[14C]acetamide into the active-site cysteine residue. Isolation of a fibrin-binding fragment expressing transglutaminase activity demonstrates that each a-chain of the dimeric Factor XIIIa could function independently to cross-link fibrin. The fibrin-binding site could play an important role in localizing Factor XIIIa to the fibrin clot.

On the Plasmin Digestion of Factor XIII

1975 ◽  
Author(s):  
K. Mikami ◽  
T. Mikami ◽  
H. Suzuki ◽  
M. Fujimaki ◽  
K. Fukutake
Keyword(s):  
Plasma Protein ◽  
Gel Electrophoresis ◽  
Factor Xiii ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Platelet Factor ◽  
Subunit A ◽  
Plasma Factor ◽  
Congenital Afibrinogenemia

The plasmin digestion of factor XIII in the plasma with congenital afibrinogenemia has been reported by Suzuki et al. in 1967. The present investigation using polyacrylamide gel electrophoresis and crossimmunoelectrophoresis with anti-factor XIII (A & S) serum demonstrates that the process of plasmin digestion of factor XIII can be divided into three steps ; the subunit A of factor XIII such as placental or platelet factor XIII is ready to get plasmin digestion following the decrease of amount and activity of subunit A, and the subunit S in the A1 S complex like plasma factor XIII is more readily affectable on the plasmin digestion than the subunit A of the complex, and the A2 S complex coexisting with fibrinogen or plasma protein is fairly stable on exposure to plasmin.

scholarly journals An investigation into the role of coagulation factor XIII in ADP- induced aggregation and fibrinogen binding with rabbit platelets

Blood ◽  
1982 ◽  
Vol 60 (4) ◽  
pp. 905-911
Author(s):  
EJ Harfenist ◽  
G Raychaudhuri ◽  
MA Packham ◽  
JF Mustard
Keyword(s):  
Factor Xiii ◽  
Specific Binding ◽  
Coagulation Factor ◽  
Factor Xiiia ◽  
Platelet Protein ◽  
Fibrinogen Binding ◽  
Plasma Factor ◽  
Rabbit Platelets ◽  
Platelet Aggregate ◽  
Induced Aggregation

Because there was a possibility that activated factor XIII (factor XIIIa) might stabilize a platelet-fibrinogen aggregate through its crosslinking action, we have isolated plasma factor XIII, activated it, and studied the effect of factor XIIIa at a concentration of 3.3 micrograms/ml on aggregation and 125I-fibrinogen binding of rabbit platelets stimulated with 9 microM ADP. Factor XIIIa did not cause aggregation in the absence of ADP, nor did it enhance ADP-induced aggregation or substantially stabilize the platelet aggregate. The presence of factor XIIIa did not affect the amount of fibrinogen bound to platelets immediately after stimulation with ADP, but it appeared to cause a slow specific binding of 125I-fibrinogen to platelets whether or not they were stimulated with ADP. This binding, which was not inhibited by prostaglandin E1, did not lead to aggregation and was accompanied by crosslinking of fibrinogen through its A alpha and gamma chains, either to other fibrinogen molecules or to a platelet protein or proteins.

scholarly journals The isolation of human platelet factor V [published erratum appears in Blood 1987 Jul;70(1):339]

Blood ◽  
1987 ◽  
Vol 69 (4) ◽  
pp. 1188-1195 ◽  
Author(s):  
RW Viskup ◽  
PB Tracy ◽  
KG Mann
Keyword(s):  
Human Plasma ◽  
Human Platelet ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Factor V ◽  
Single Chain ◽  
Platelet Factor ◽  
Factor Va ◽  
Plasma Factor ◽  
End Products

Abstract Human platelet factor V has been isolated using either a monoclonal or polyclonal antibody directed against human plasma factor V. The largest peptide observed upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of purified human platelet factor V comigrates with purified human plasma factor V. However, a significant portion of the isolated protein is represented by peptides of lower apparent molecular weight (Mr). These lower Mr species that copurify with platelet factor V have been shown to be platelet factor V components by their immunological cross-reactivity with monoclonal and polyclonal antibodies to purified human plasma factor V. Platelets isolated from whole blood drawn directly into inhibitors to prevent proteolysis and platelet activation demonstrate the pattern of fragmented platelet factor V. The components of purified platelet factor V demonstrate apparent Mr ranging between 115 K and 330 K and are detectably different from the intermediates and end products observed during the thrombin cleavage of single-chain plasma factor V. Upon treatment with thrombin the platelet factor V components are cleaved and the end products are indistinguishable from those obtained upon thrombin activation of plasma factor V to plasma factor Va. Examination of the components by immunoblotting demonstrates that some of the cleavages which have occurred in the platelet factor V molecule are within the 150-K activation peptide. Bioassay indicates that platelet factor V exists as a procofactor and cleavage by thrombin yields the active cofactor, platelet factor Va. These data suggest that human platelet factor V is stored in the platelet as a partially fragmented procofactor that can be activated by thrombin to yield human platelet factor Va, the active cofactor in the human prothrombinase complex.

scholarly journals An autosomal dominant, qualitative platelet disorder associated with multimerin deficiency, abnormalities in platelet factor V, thrombospondin, von Willebrand factor, and fibrinogen and an epinephrine aggregation defect

Blood ◽  
1996 ◽  
Vol 87 (12) ◽  
pp. 4967-4978 ◽  
Author(s):  
CP Hayward ◽  
GE Rivard ◽  
WH Kane ◽  
J Drouin ◽  
S Zheng ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Factor V ◽  
Binding Studies ◽  
Platelet Factor ◽  
Platelet Disorder ◽  
Plasma Factor ◽  
Von Willebrand ◽  
Willebrand Factor

Multimerin is a massive soluble, multimeric protein found in platelets and endothelial cells. Recent studies identified multimerin as a specific coagulation factor V binding protein, complexed with platelet, but not plasma, factor V. These findings led us to investigate individuals with inherited factor V deficiencies for possible multimerin abnormalities. Platelet proteins were evaluated using immunoassays, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting, immunoprecipitation, and direct binding studies. Patients with factor V Quebec, a disorder with abnormal platelet factor V, had a quantitative deficiency in multimerin (n = 11 tested; mean, 12.5%; range, 5% to 27% of the normal pool; normal range, 45% to 214%) with a normal multimer pattern. Quantitative and qualitative abnormalities were detected in their platelet factor V. An unrelated patient who was deficient in platelet and plasma factor V had normal platelet multimerin. The levels of platelet beta- thromboglobulin, von Willebrand factor, thrombospondin, and fibrinogen antigen were normal in the factor V Quebec patients. However, proteins with abnormal mobility were detected in their platelet lysate and releasate, and their platelet thrombospondin, von Willebrand factor, and fibrinogen showed evidence of proteolytic degradation. Platelet counts of the factor V Quebec patients ranged from mildly thrombocytopenic to low normal (mean, 159 x 10(9)/L; range, 104 to 198 x 10(9)/L). In addition, their platelets failed to aggregate in response to 6 to 10 micromol/L epinephrine despite normal numbers of platelet alpha 2-adrenergic receptors. These data indicate that patients with factor V Quebec have an inherited bleeding disorder distinct from other platelet disorders and associated with multiple abnormalities, including multimerin deficiency, abnormal platelet factor V, thrombospondin, von Willebrand factor, and fibrinogen, and an epinephrine aggregation defect.

scholarly journals The isolation of human platelet factor V [published erratum appears in Blood 1987 Jul;70(1):339]

Blood ◽  
1987 ◽  
Vol 69 (4) ◽  
pp. 1188-1195 ◽  
Author(s):  
RW Viskup ◽  
PB Tracy ◽  
KG Mann
Keyword(s):  
Human Plasma ◽  
Human Platelet ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Factor V ◽  
Single Chain ◽  
Platelet Factor ◽  
Factor Va ◽  
Plasma Factor ◽  
End Products

Human platelet factor V has been isolated using either a monoclonal or polyclonal antibody directed against human plasma factor V. The largest peptide observed upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of purified human platelet factor V comigrates with purified human plasma factor V. However, a significant portion of the isolated protein is represented by peptides of lower apparent molecular weight (Mr). These lower Mr species that copurify with platelet factor V have been shown to be platelet factor V components by their immunological cross-reactivity with monoclonal and polyclonal antibodies to purified human plasma factor V. Platelets isolated from whole blood drawn directly into inhibitors to prevent proteolysis and platelet activation demonstrate the pattern of fragmented platelet factor V. The components of purified platelet factor V demonstrate apparent Mr ranging between 115 K and 330 K and are detectably different from the intermediates and end products observed during the thrombin cleavage of single-chain plasma factor V. Upon treatment with thrombin the platelet factor V components are cleaved and the end products are indistinguishable from those obtained upon thrombin activation of plasma factor V to plasma factor Va. Examination of the components by immunoblotting demonstrates that some of the cleavages which have occurred in the platelet factor V molecule are within the 150-K activation peptide. Bioassay indicates that platelet factor V exists as a procofactor and cleavage by thrombin yields the active cofactor, platelet factor Va. These data suggest that human platelet factor V is stored in the platelet as a partially fragmented procofactor that can be activated by thrombin to yield human platelet factor Va, the active cofactor in the human prothrombinase complex.

Enhanced Affinity of Fibrinogen and Factor XIII Induced by Limited Proteolysis

1979 ◽  
Author(s):  
S.I. Chung ◽  
J.E. Folk ◽  
J.S. Finlayson
Keyword(s):  
Form Factor ◽  
Affinity Chromatography ◽  
Factor Xiii ◽  
Tissue Transglutaminase ◽  
Limited Proteolysis ◽  
Factor Xiiia ◽  
Salt Precipitation ◽  
Salting Out ◽  
Platelet Factor ◽  
Plasma Factor

Previous studies have shown that tissue transglutaminase has strong affinity for fibrinogen. In this study, the binding of protransglutaminase, i.e., plasma Factor XIII(a2b2) and platelet Factor XIII (a2), to fibrinogen was examined by ultracentrifugation, exclusion and affinity chromatography, Immunoelectrophoresis, and salt precipitation. Affinity was detected only by salting out and affinity chromatography. Limited proteolysis to form Factor XIIIa (a′2b2 and a′2) enhanced the affinity for fibrinogen. Limited proteolysis of fibrinogen by thrombin or ancrod enhanced affinity for both a2b2 and a2. Fibrin also bound a′2b2 and a′2. The b2 subunit exhibited no affinity for fibrinogen or fibrin. Thus, in the presence of fibrin(ogen), a′2b2 gave rise to a fibrin(ogen)-a′2 complex, and the b2 subunit was liberated.The enhanced affinity for fibrinogen induced by limited proteolysis of Factor XIII suggests that fibrinogen could serve to adsorb active transglutaminase(s) released or generated in the circulation.

scholarly journals An investigation into the role of coagulation factor XIII in ADP- induced aggregation and fibrinogen binding with rabbit platelets

Blood ◽  
1982 ◽  
Vol 60 (4) ◽  
pp. 905-911 ◽  
Author(s):  
EJ Harfenist ◽  
G Raychaudhuri ◽  
MA Packham ◽  
JF Mustard
Keyword(s):  
Factor Xiii ◽  
Specific Binding ◽  
Coagulation Factor ◽  
Factor Xiiia ◽  
Platelet Protein ◽  
Fibrinogen Binding ◽  
Plasma Factor ◽  
Rabbit Platelets ◽  
Platelet Aggregate ◽  
Induced Aggregation

Abstract Because there was a possibility that activated factor XIII (factor XIIIa) might stabilize a platelet-fibrinogen aggregate through its crosslinking action, we have isolated plasma factor XIII, activated it, and studied the effect of factor XIIIa at a concentration of 3.3 micrograms/ml on aggregation and 125I-fibrinogen binding of rabbit platelets stimulated with 9 microM ADP. Factor XIIIa did not cause aggregation in the absence of ADP, nor did it enhance ADP-induced aggregation or substantially stabilize the platelet aggregate. The presence of factor XIIIa did not affect the amount of fibrinogen bound to platelets immediately after stimulation with ADP, but it appeared to cause a slow specific binding of 125I-fibrinogen to platelets whether or not they were stimulated with ADP. This binding, which was not inhibited by prostaglandin E1, did not lead to aggregation and was accompanied by crosslinking of fibrinogen through its A alpha and gamma chains, either to other fibrinogen molecules or to a platelet protein or proteins.

Plasma Factor XIII and Platelet Factor XIII in Hyperlipaemia

1976 ◽  
Vol 36 (03) ◽  
pp. 542-550 ◽  
Author(s):  
Mircea P. Cucuianu ◽  
K Miloszewski ◽  
D Porutiu ◽  
M. S Losowsky
Keyword(s):  
Factor Xiii ◽  
Significant Contribution ◽  
Quantitative Assay ◽  
Factor Xii ◽  
Platelet Factor ◽  
Type Iv ◽  
Plasma Factor ◽  
Hepatic Synthesis

SummaryPlasma factor XIII activity measured by a quantitative assay was found to be significantly higher in hypertriglyceridaemic patients (type IV and combined hyperlipoproteinaemia), as compared to normolipaemic controls. No such elevation in plasma factor XIII activity was found in patients with type IIa hyperlipaemia. Plasma pseudocholinesterase was found to parallel the elevated factor XIII activity in hypertriglyceridaemic subjects.In contrast, platelet factor XIII activity was not raised in hyperlipaemic subjects, and plasma factor XIII was found to be normal in a normolipaemic subject with throm-bocythaemia.It was concluded that there is no significant contribution from platelets to plasma factor XIII activity, and that the observed increase in plasma factor XII in hypertriglyceridaemia results from enhanced hepatic synthesis of the enzyme.

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