scholarly journals Non-proteolytic activation of cellular protransglutaminase (placenta macrophage factor XIII)

1990 ◽  
Vol 267 (2) ◽  
pp. 557-560 ◽  
Author(s):  
J Polgár ◽  
V Hidasi ◽  
L Muszbek
Keyword(s):  
Factor Xiii ◽  
Specific Activity ◽  
Activation Process ◽  
Proteolytic Activation ◽  
Plasma Factor ◽  
Nacl Concentration ◽  
Catalytically Active ◽  
Thrombin Activation ◽  
A Subunit ◽  
High Concentrations

Plasma Factor XIII is a zymogen (plasma protransglutaminase) with the tetrametric structure A2B2, whereas the cellular protransglutaminase, i.e. Factor XIII in the platelet and monocyte/macrophage, consists exclusively of A subunits (A2). It is generally accepted that at Ca2+ concentrations comparable with that in plasma the proteolytic removal of an N-terminal activation peptide is the prerequisite for the Ca2(+)-induced formation of a catalytically active configuration of subunit A. In this study it was demonstrated that at high concentrations NaCl or KCl induced a non-proteolytic activation of cellular (placental macrophage) but not plasma protransglutaminase. The activation depended on time and salt concentration, and Ca2+, in the range 0-20 mM, greatly enhanced the activation process. At 1.25 M-NaCl maximal activation occurred within 60 min in the presence of 2 mM-CaCl2, and even at physiological NaCl concentration a slow progressive activation could be observed in the presence of Ca2+. The specific activity of salt-activated Factor XIII was 1.5-2.0-fold higher than that obtained after thrombin activation. The non-proteolytic activation of cellular protransglutaminase was abolished by the addition of subunit B of plasma Factor XIII in stoichiometric amount, which suggests that (one of) the physiological function(s) of the B subunit in plasma Factor XIII is to prevent the slow spontaneous activation of A subunit that would occur in a plasmatic environment.

scholarly journals Val34Leu polymorphism of plasma factor XIII: biochemistry and epidemiology in familial thrombophilia

Blood ◽  
2000 ◽  
Vol 96 (7) ◽  
pp. 2479-2486 ◽  
Author(s):  
István Balogh ◽  
Gabriella Szôke ◽  
Levente Kárpáti ◽  
Ulla Wartiovaara ◽  
Éva Katona ◽  
...  
Keyword(s):  
Protective Effect ◽  
Venous Thrombosis ◽  
Factor Xiii ◽  
Coagulation Factor ◽  
Wild Type ◽  
Thrombin Cleavage ◽  
Plasma Factor ◽  
Thrombin Activation ◽  
A Subunit ◽  
The Mean

Abstract Val34Leu polymorphism of the A subunit of coagulation factor XIII (FXIII-A) is located in the activation peptide (AP) just 3 amino acids away from the thrombin cleavage site. This mutation has been associated with a protective effect against occlusive arterial diseases and venous thrombosis; however, its biochemical consequences have not been explored. In the current study it was demonstrated that the intracellular stability and the plasma concentration of FXIII of different Val34Leu genotypes are identical, which suggests that there is no difference in the rate of synthesis and externalization of wild-type and mutant FXIII-A. In contrast, the release of AP by thrombin from the Leu34 allele proceeded significantly faster than from its wild-type Val34 counterpart. By molecular modeling larger interaction energy was calculated between the Leu34 variant and the respective domains of thrombin than between the Val34 variant and thrombin. In agreement with these findings, the activation of mutant plasma FXIII by thrombin was faster and required less thrombin than that of the wild-type variant. Full thrombin activation of purified plasma FXIII of different genotypes, however, resulted in identical specific transglutaminase activities. Similarly, the mean specific FXIII activity in the plasma was the same in the groups with wild-type, heterozygous, and homozygous variants. Faster activation of the Leu34 allele hardly could be associated with its presumed protective effect against venous thrombosis. No such protective effect was observed in a large group of patients with familial thrombophilia.

PLASMA FACTOR XIII AND ACTIVATION PROCESS ANALYZED BY ELISA METHOD FOR A2B2 COMPLEX.

1987 ◽  
Author(s):  
K Koike ◽  
M Hada ◽  
H Yorifuji ◽  
S Ikematsu ◽  
K Fukutake ◽  
...  
Keyword(s):  
Calcium Ion ◽  
Factor Xiii ◽  
Molecular Conformation ◽  
Fibrin Clot ◽  
Activation Process ◽  
Measurement Sensitivity ◽  
B Subunit ◽  
Plasma Factor ◽  
A Subunit ◽  
Subunit B

Factor XIII induces the crosslinking of fibrin at terminal stage of blood coagulation. New ELISA methods of a-subunit, b-subunit and a2b2 complex of factor XIII were developed by the author and the following results concerning the activation process of factor XIII were obtained. New ELISA methods for a-subunit, b-subunit and a2b2 complex of factor XIII were specific with high sensitivities on each items and indicated the measurement capacity for simultaneous quantitation of many samples, more over we looked upon the dissociation of a2b2 complex with these methods to able to analyze at the subunit levels of factor XIII in details. When a-subunit dimer of platelets was discharged into the plasma by the use of freezing and thawing method on PRP, it was more easily affected to lose their antigeniety than that of a2b2 complex, a-subunit levels in the plasma of congenital factor XIII deficiencies were measured in very low concentration or below the measurement sensitivity of this ELISA, b-subunit levels in the same plasma were indicated around the half of normal levels. These results were as same as another immunological method. It was suggested that the molecular conformation of a-subunit could be changed by the addition of thrombin in high concentration and consequently a-subunit with thrombin was modifyed to show high antigeniety. It was observed that a2b2 complex was dissociated by the addition of thrombin without calcium ion, and the process of this dissociation of a2b2 complex was remarkably accelerated by the addition of calcium ion. Because a-subunit and b-subunit were adsorved on fibrin clot, it might be presumed that the crosslinking of fibrin molecules could be accelerated locally on the surface of fibrin clot.

scholarly journals Val34Leu polymorphism of plasma factor XIII: biochemistry and epidemiology in familial thrombophilia

Blood ◽  
2000 ◽  
Vol 96 (7) ◽  
pp. 2479-2486 ◽  
Author(s):  
István Balogh ◽  
Gabriella Szôke ◽  
Levente Kárpáti ◽  
Ulla Wartiovaara ◽  
Éva Katona ◽  
...  
Keyword(s):  
Protective Effect ◽  
Venous Thrombosis ◽  
Factor Xiii ◽  
Coagulation Factor ◽  
Wild Type ◽  
Thrombin Cleavage ◽  
Plasma Factor ◽  
Thrombin Activation ◽  
A Subunit ◽  
The Mean

Val34Leu polymorphism of the A subunit of coagulation factor XIII (FXIII-A) is located in the activation peptide (AP) just 3 amino acids away from the thrombin cleavage site. This mutation has been associated with a protective effect against occlusive arterial diseases and venous thrombosis; however, its biochemical consequences have not been explored. In the current study it was demonstrated that the intracellular stability and the plasma concentration of FXIII of different Val34Leu genotypes are identical, which suggests that there is no difference in the rate of synthesis and externalization of wild-type and mutant FXIII-A. In contrast, the release of AP by thrombin from the Leu34 allele proceeded significantly faster than from its wild-type Val34 counterpart. By molecular modeling larger interaction energy was calculated between the Leu34 variant and the respective domains of thrombin than between the Val34 variant and thrombin. In agreement with these findings, the activation of mutant plasma FXIII by thrombin was faster and required less thrombin than that of the wild-type variant. Full thrombin activation of purified plasma FXIII of different genotypes, however, resulted in identical specific transglutaminase activities. Similarly, the mean specific FXIII activity in the plasma was the same in the groups with wild-type, heterozygous, and homozygous variants. Faster activation of the Leu34 allele hardly could be associated with its presumed protective effect against venous thrombosis. No such protective effect was observed in a large group of patients with familial thrombophilia.

scholarly journals The action of o-dihydric phenols in the hydroxylation of p-coumaric acid by a phenolase from leaves of spinach beet (Beta vulgaris L.)

1970 ◽  
Vol 119 (1) ◽  
pp. 89-94 ◽  
Author(s):  
P. F. T. Vaughan ◽  
V. S. Butt
Keyword(s):  
Caffeic Acid ◽  
Beta Vulgaris ◽  
Oxidase Activity ◽  
Maximum Rate ◽  
Specific Activity ◽  
Catechol Oxidase ◽  
Coumaric Acid ◽  
Catalytically Active ◽  
Lag Period ◽  
High Concentrations

1. Under defined conditions, the hydroxylation of p-coumaric acid catalysed by a phenolase from leaves of spinach beet (Beta vulgaris L.) was observed to develop its maximum rate only after a lag period. 2. By decreasing the reaction rate with lower enzyme concentrations or by increasing it with higher concentrations of reductants, the length of the lag period was inversely related to the maximum rate subsequently developed. 3. Low concentrations of caffeic acid or other o-dihydric phenols abolished this lag period. With caffeic acid, the rate of hydroxylation was independent of the reductant employed. 4. Hydroxylation was inhibited by diethyldithiocarbamate, but with low inhibitor concentrations hydroxylation recovered after a lag period. This lag could again be abolished by the addition of high concentrations of caffeic acid or other o-dihydric phenols. 5. Catechol oxidase activity showed no lag period, and did not recover from diethyldithiocarbamate inhibition. 6. The purified enzyme contained 0.17–0.33% copper; preparations with the highest specific activity were found to have the highest copper content. 7. The results are interpreted to suggest that the oxidation of o-dihydric phenols converts the enzymic copper into a species catalytically active in hydroxylation. This may represent the primary function for the catechol oxidase activity of the phenolase complex. The electron donors are concerned mainly, but not entirely, in the reduction of o-quinones produced in this reaction.

Association of a Common Polymorphism in the Factor XIII Gene With Venous Thrombosis

Blood ◽  
1999 ◽  
Vol 93 (3) ◽  
pp. 906-908 ◽  
Author(s):  
Andrew J. Catto ◽  
Hans P. Kohler ◽  
Julie Coore ◽  
Michael W. Mansfield ◽  
Max H. Stickland ◽  
...  
Keyword(s):  
Venous Thrombosis ◽  
Factor Xiii ◽  
Factor V Leiden ◽  
Common Mutation ◽  
Factor V ◽  
Site Factor ◽  
Gene Coding ◽  
History Of ◽  
Thrombin Activation ◽  
A Subunit

We have shown an association between a common mutation in the factor XIII a-subunit gene, coding for an amino acid change, 3 amino acids from the thrombin activation site (factor XIII Val34Leu) that may protect against myocardial infarction and predisposes to intracranial hemorrhage. To investigate the possible role of factor XIII Val34Leu in the pathogenesis of venous thromboembolism (VTE) and potential interactions with factor V Leiden (FV:Q506) and prothrombin G → A 20210, we studied 221 patients with a history of VTE and 254 healthy controls. Patients with VTE showed an increased frequency of the FXIII Val/Val genotype (63% v 49%) and a lower frequency of the Val/Leu genotype (31% v 42%) than controls (P = .007). FV:Q506 heterozygotes were more frequent in VTE patients (11%) than controls (5%; P = .04). The prothrombin G → A 20210 mutation was present in only 3 patients and no controls (P = .10). In a logistic regression model for a history of VTE, the odds ratio (95% confidence interval) for FXIII Val/Leu or Leu/Leu genotype was 0.63 (0.38 to 0.82) and for possession of FV:Q506 2.40 (1.17 to 4.90). There was no evidence for an interaction between factor XIII Val34Leu genotype and FV:Q506, prothrombin G → A 20210, sex, or age. It is concluded that possession of the Leu allele at factor XIII Val34Leu is protective against deep venous thrombosis.

scholarly journals Factor V turnover in a primate model

Blood ◽  
1995 ◽  
Vol 86 (7) ◽  
pp. 2616-2623 ◽  
Author(s):  
MD Rand ◽  
SR Hanson ◽  
KG Mann
Keyword(s):  
Specific Activity ◽  
Apparent Molecular Weight ◽  
In Vivo Model ◽  
Normal Male ◽  
Factor V ◽  
Primate Model ◽  
Isolation And Characterization ◽  
Plasma Factor ◽  
Thrombin Activation

The isolation and characterization of baboon plasma factor V (FV) were performed for the development of an in vivo model for studying factor V/Va physiology in nonhuman primates. Baboon FV was purified by immunoaffinity chromatography with an antihuman FV monoclonal antibody and exhibits a specific activity of 1,940 U/mg. Baboon FV activation by thrombin proceeds through two proteolytic pathways similar to those observed with human and bovine FV. Limited amino acid sequencing of FV and its thrombin activation fragments shows 95% identity with human and 79% identity with bovine FV. 125I-Factor V and a mixture of thrombin cleaved 125I-FV activation products were infused into normal male baboons and evaluated by blood sample radioactivity measurements and by autoradiography of plasma samples following resolution by gel electrophoresis. Factor V disappeared with a half-life (t1/2) of 12.98 +/- 1.85 hours and was cleared without obvious degradation of the molecule during circulation. The radioactivity associated with the thrombin activated FV mixture, which consisted of the Mr = 220,000 activation intermediate, the Mr = 150,000 activation peptide, the heavy chain (HC) and the light chain (LC) of FVa, was cleared in a nonlinear manner. The HC and LC were removed with t1/2 < 20 minutes. The apparent molecular weight (Mr) = 220,000 and Mr = 150,000 fragments were cleared with t1/2 > 6 hours and t1/2 > 30 hours, respectively.

scholarly journals Detection of factor XIIIa (active fibrin-stabilizing factor) in normal plasma

Blood ◽  
1978 ◽  
Vol 52 (3) ◽  
pp. 581-591 ◽  
Author(s):  
JC Nelson ◽  
RG Lerner
Keyword(s):  
Factor Xiii ◽  
Specific Activity ◽  
Proteolytic Enzymes ◽  
High Specific Activity ◽  
Factor Xa ◽  
Factor Xiiia ◽  
Calcium Dependent ◽  
Normal Plasma ◽  
Plasma Factor

Abstract Factor XIIIa (active fibrin-stabilizing factor) generated in heat- defibrinated plasma by the addition of thrombin can be measured by 14C- putrescine incorporation into casein. Modification of this assay be substituting 3H-putrescine of high specific activity as the donor amine permits measurement of amine incorporation by plasma even in the absence of added thrombin. Incorporation is calcium dependent, inhibited by iodoacetamide, and absent from congenital factor XIII- deficient plasma and from normal platelets. The transamidating activity detected by radioenzymatic assay catalyzed the formation of gamma-gamma dimers and alpha polymers of fibrin and was thus biologically functional. This fibrin cross-linking activity was absent from factor XIII-deficient plasma. These experiments show (1) some factor XIII is present in plasma as factor XIIIa; (2) this factor XIIIa can cross-link fibrin and thus has biologic activity as well; and (3) this activity is not present in factor XIII-deficient plasma. Factor XIIIa in normal plasma is possibly activated in vivo, perhaps by circulating thrombin, factor Xa, or other proteolytic enzymes.

The Effect Of Plasmin On Factor XIII (Fibrin Stabilizing Factor)

1981 ◽  
Author(s):  
D M Rider ◽  
J M McDonagh
Keyword(s):  
Factor Xiii ◽  
Activate Factor ◽  
Polyacrylamide Gels ◽  
Clotting Factors ◽  
Human Fibrinogen ◽  
Platelet Factor ◽  
Plasma Factor ◽  
Before And After ◽  
A Subunit ◽  
Almost All

The action of plasmin on several blood clotting factors has been studied; however, controversy exists concerning the effect of plasmin on factor XIII. Factor XIII was purified from plasma and platelets and then exposed to plasmin for up to 6 hours. Plasmin to factor XIII ratios ranged from 0.03-0.1 casein units plasmin per mg factor XIII. These plasmin levels exhibited strong proteolytic activities against B-casein and purified human fibrinogen Following incubation of factor XIII (activated and unactivated) with plasmin the mixtures were electrophoresed on 7% SDS-polyacrylamide gels. The factor XIII preparations were assayed for 14C-putrescine incorporating activity before and after exposure to plasmin. Platelet factor XIII was,labeled With 125Iodine and lableled a subunit (activated and unactivated) was exposed to plalmin for up to 2 hours. These mixtures were electrophoresed on 12.5% Urea-SDS Polyacrylamide gels and a radioactivity profile was determined for each gel.Following extensive exposure to Plasmin the relative molecular weights of the factor XIII subunits (a, a* and b)remained constant and almost all (90-100%) of the 14C-put-rescine incorporating activity was recovered. The radio-activity profiles of the gels of 125I-labeled platelet factor XIII were identical before and after incubation with plasmin. Plasmin did not activate factor XIII in the assay system nor did factor XIII inactivate plasmin by crosslinking it. These experiments indicate that plasmin does not activate or degrade factor XIII and that the b subunit of plasma factor XIII plays no role in protecting the a subunit from the action of plasmin.

Association of a Common Polymorphism in the Factor XIII Gene with Myocardial Infarction

1998 ◽  
Vol 79 (01) ◽  
pp. 8-13 ◽  
Author(s):  
Hans Kohler ◽  
Max Stickland ◽  
Nicholas Ossei-Gerning ◽  
Angela Carter ◽  
Hanna Mikkola ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Factor Xiii ◽  
Exon 2 ◽  
Site Factor ◽  
Genotype Frequencies ◽  
History Of ◽  
Thrombin Activation ◽  
A Subunit ◽  
Artery Disease ◽  
Pai 1

SummaryFactor XIII when activated by thrombin, crosslinks fibrin, however its role in thrombotic disorders is unknown. A common point mutation (G T) in exon 2 of the A-subunit gene which codes for an amino acid change three amino acids from the thrombin activation site (Factor XIIIVal34Leu) is a candidate for a role in the pathogenesis of acute myocardial infarction. Factor XIII genotype frequencies were determined in a case-control study of 398 caucasian patients and 196 healthy controls. Patients had undergone angiography for investigation of coronary artery disease and were evaluated for a history of myocardial infarction. The prevalence of the mutation was lower in patients with myocardial infarction than without (32% vs. 50%), p = 0.0009 and than in controls (32% vs. 48%), p = 0.005. Patients possessing the mutation with a history of myocardial infarction had higher PAI-1 concentrations (mean, 27.9 vs. 16.7 ng/ml, p = 0.004) and the PAI-1 4G/4G genotype was commoner (43% vs. 26%, p = 0.03). There was no difference in PAI-1 4G/4G genotype (33% vs. 32%) and PAI-1 levels (mean, 21.0 vs. 20.9 ng/ml) in patients possessing wild type with MI compared to those without MI. These results indicate that the G T mutation coding for factor XIIIVal34Leu is protective against myocardial infarction and suggest a mechanism whereby elevated levels of PAI-1 may contribute to vascular risk.

THROMBOMODULIN INHIBITS THE ACTIVATION OF FACTOR XIII BY THROMBIN.

1987 ◽  
Author(s):  
J Polgar ◽  
I Lerant ◽  
L Muszbek ◽  
R Machovich
Keyword(s):  
Endothelial Cell ◽  
Factor Xiii ◽  
Protein C ◽  
Receptor Protein ◽  
Activation Process ◽  
Factor V ◽  
Kinetic Assay ◽  
Active Structure ◽  
Significant Difference ◽  
A Subunit

The thrombogenic functions of thrombin, studied so far, are diminished or blocked when thrombin is bound to the endothelial cell via its receptor protein thrombomodulin. The thrombomodu-lin-thrombin complex fails to clot fibrinogen, to activate platelet and Factor V, while the activation of the antithrombo-genic protein, protein C is extremely enhanced. Although the activation of Factor XIII (FXIIl) belongs to the thrombogenic functions of thrombin, the effect of thrombomodulin on this process has not been investigated, so far. The aim of this study was to establish whether the presence of thrombomodulin modifies the effect of thrombin on FXIII. The activation was followed by measuring the transglutaminase activity of FXIIIa formed using our UV-kinetic assay (Muszbek et al., Clin. Chem. , 3L, 35, 1985) and by monitoring the amount of activation peptide split off by thrombin from the a subunit. The time dependence of FXIII activation using various thrombin concentrations showed significant difference in the presence and absence of thrombomodulin. Thrombomodulin significantly slowed down the activity of thrombin toward FXIII but did not prevent it completely. The possibility that thrombomodulin influences changes brought about by Ca and noij+the action of thrombin was excluded. When thrombomodulin and Ca were added only after the proteolytic leavage of FXIII had taken place, it had no effect on the Ca induced activation process. The results suggests that thrombomodulin inhibits the rate of conversion of FXIII to its enzymatically active structure but does not influence the amount of FXIIIa formed.

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