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.

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.

Characteristics of Platelet Protransglutaminase (Factor XIII)-Binding Activity in Human Plasma

1975 ◽  
Author(s):  
D. Bannerjee ◽  
M. W. Mosesson
Keyword(s):  
Human Plasma ◽  
Ammonium Sulfate ◽  
Factor Xiii ◽  
Binding Activity ◽  
Platelet Factor ◽  
Catalytic Potential ◽  
Plasma Factor ◽  
Exclusion Chromatography ◽  
A Chain ◽  
Polypeptide Chains

Human plasma Factor XIII (F. XIII) complex is composed of two types of noncovalently linked polypeptide chains termed a and b; only the a chain possesses catalytic potential. Platelet Factor XIII is comprised solely of a chains which are identical to those found in plasma. In this study platelets were utilized as a source of unbound a chains to characterize F. XIII (a chain)-binding activity in plasma and its subfractions. Upon exclusion chromatography of unheated plasma or of an unheated ammonium sulfate (20% sat.) subfraction, F. XIII activity emerged in a peak corresponding to a mol. wt. of < 500,000 (region 1). If these samples had first been heated at 60° to precipitate fibrinogen, F. XIII was eluted in a peak corresponding to a mol wt. of about 300,000 (region 2). Chromatography of the platelet zymogen alone resulted in a F. XIII peak corresponding in position to that of monomeric a chain (mol. wt. 80,000, region 3).Exclusion chromatography of the unheated ammonium sulfate fraction yielded, in addition to the F. XIII peak in region 1, a protein peak (peak II) in region 2 which contained no F. XIII. When peak II was mixed with platelet F. XIII, and again subjected to exclusion chromatography, the platelet F. XIII peak shifted from its expected position in region 3 and emerged instead in region 2 ; this behavior demonstrated F. XIII (a chain)-binding activity within peak II. The same chromatographic shift was observed in mixtures of platelet F. XIII and normal plasma or that from a patient with congenital F. XIII (a chain) deficiency. Immunochemical analyses of chromatographic fractions indicated that a chain-binding was due to complexing of a chains with freely circulating b chains. Since a chains and b chains have different biosynthetic sites we conclude that b chains serve as an extracellular F. XIII (a chain)-binding protein.Supported by NHLI grant HL-11409.

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.

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.

Platelet Factor XIII Becomes Active without the Release of Activation Peptide during Platelet Activation

1993 ◽  
Vol 69 (03) ◽  
pp. 282-285 ◽  
Author(s):  
László Muszbek ◽  
János Polgár ◽  
Zoltán Boda
Keyword(s):  
Platelet Activation ◽  
Factor Xiii ◽  
Platelet Factor ◽  
Subunit A ◽  
Cellular Factor ◽  
B Subunit ◽  
Exact Function ◽  
Plasma Factor ◽  
A Subunit ◽  
Activation Peptide

SummaryThe potentially active A subunit of factor XIII of blood coagulation has also been detected in platelets and monocytes/macrophages though the exact function of this cellular protransglutaminase has not yet been elucidated. In physiological conditions the first step in the activation of plasma factor XIII is the removal of an activation peptide from the N-terminal end of subunit A by thrombin. The A subunit then, in the presence of Ca2+, dissociates from the inhibitory B subunit and assumes an active conformation. Cellular factor XIII, which lacks B subunit, can be proteolytically activated in vitro by thrombin and the intracellular Ca2+ sensitive protease, calpain, in the same way as plasma factor XIII subunit A, and calpain has been suggested as the intracellular protease involved in the activation of cellular factor XIII in platelets. In the present experiments it was shown by SDS PAGE that during long-term stimulation of platelets with thrombin nondisulfide-crosslinked high M r protein polymers not penetrating the concentrating gel were formed. The lack of these polymers in thrombin-stimulated factor XIII deficient platelets clearly indicated that their formation in normal platelets was due to factor XIII that became active during platelet activation. However, no release of the activation peptide could be detected by Western blotting during this process. Similarly, no proteolytic cleavage of factor XIII was detectable when platelets were stimulated by Ca2+ ionophore through this stimulus activated calpain as it was clearly demonstrated by the breakdown of major intracellular calpain substrates. The results indicate: 1) during thrombin induced platelet activation factor XIII becomes active and crosslinks platelet protein, 2) platelet factor XIII is not an intracellular substrate of calpain, 3) cellular factor XIII could be activated without the proteolytic removal of activation peptide. It is presumed that the nonproteolytic pathway for the activation of cellular factor XIII, we reported most recently, might have physiological implications under such conditions.

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.

The Effect of Major Surgical Procedures on Plasma and Platelet Levels of Factor XIII

1974 ◽  
Vol 31 (01) ◽  
pp. 020-029 ◽  
Author(s):  
Beth A Letheby ◽  
Richard B Davis ◽  
Arden E Larsen
Keyword(s):  
Blood Coagulation ◽  
Surgical Procedures ◽  
Orthopedic Surgery ◽  
Factor Xiii ◽  
Normal Blood ◽  
Neoplastic Disease ◽  
Platelet Factor ◽  
Linear Decrease ◽  
Plasma Factor

SummaryFactor XIII (FSF or fibrinoligase), an enzyme required for normal blood coagulation, can be measured by its catalysis of the incorporation of a fluorescent amine, monodansylcadaverine (N-(5-aminopentyl)-5-dimethylamino-l-naphthalene-sulfon-amide) into casein. The amine incorporation test was applied to fourteen patients who had major operations. Levels of factor XIII were measured before surgery and at intervals of two, four, and six days after operation. Plasma factor XIII showed a significant, linear decrease for each day after surgery. Patients having orthopedic surgery showed less pronounced changes than the remaining patients. Factor XIII also decreased after operation in patients with neoplastic disease, but the change was not different from that after general surgical procedures done in the absence of neoplasia. Platelet factor XIII levels were increased on the second day after operation, diminished on day four, and increased above pre-operative levels on the sixth day after surgery. Possible explanations of the observations are presented.

Interactions of Factor XIII Subunits as Studied by Affinity Chromatography

1975 ◽  
Author(s):  
Jan McDonagh ◽  
Richard P. McDonagh
Keyword(s):  
Ionic Strength ◽  
Affinity Chromatography ◽  
Factor Xiii ◽  
National Institutes Of Health ◽  
Platelet Factor ◽  
B Subunit ◽  
Plasma Factor ◽  
A Subunit ◽  
Low Ionic Strength ◽  
Native Plasma

A new method for studying the interactions of plasma factor XIII subunits, as well as purifying both plasma and platelet factor XIII, has been developed using an organomer-curial linked to agarose. Native plasma factor XIII a subunit binds to the ligand accompanied by b subunit, which can be dissociated from the ligated a subunit by treatment with thrombin and calcium. With various combinations of thrombin, calcium, and alteration of ionic strength it has been shown that: (a) both unactivated a subunit and thrombin activated a’ subunit bind to the ligand; (b) “excess” b subunit present in highly purified preparations of plasma factor XIII dissociates and can be eluted at low ionic strength in the absence of calcium or thrombin; (c) in our preparations of highly purified plasma factor XIII approximately 40% of b subunit is in “excess”; (d) the remaining b subunit can be eluted with thrombin and calcium; (e) both a and a’ subunits can be eluted with dithiothreitol ; (f ) when plasma factor XIII zymogen is applied to the column approximately equal amounts of a and b subunits co-elute with dithiothreitol. These observations offer further evidence for the roles of thrombin and calcium in the activation of plasma factor XIII and provide a new tool for studying the structure-function relationships of factor XIII.Supported by Grants HL 14228 and HL 06350 from the National Institutes of Health, U.S.A.

scholarly journals The Involvement of Factors XII & XI in the Platelet Coagulant Activity

1977 ◽  
Author(s):  
J.M. Connellan ◽  
S. Bowden ◽  
P.A. Castaldi
Keyword(s):  
Platelet Rich Plasma ◽  
Factor Xii ◽  
Platelet Activity ◽  
Freezing And Thawing ◽  
Clotting Factors ◽  
Factor Xi ◽  
Platelet Factor ◽  
Coagulant Activity ◽  
Plasma Factor ◽  
Rapid Purification

Platelets provide coagulant activities, other than the Phospholipid Platelet factor 3, which can be released from the platelet. In Platelet Poor Plasma, the initiation of the intrinsic pathway is proposed to proceed via factors Xll & XI as well as 2 other factors, Prekallekrein and Kininogen. The role of these factors in the initiation of coagulation in Platelet Rich Plasma is under investigation. Walsh, (Br.J. Haemat. 22. 393, 1972) and Muntz (Haemostasis in press 1977), have shown that platelets devoid of factor Xll will still produce a coagulant activity when Platelet Rich Plasma is incubated with collagen. However, we have recently shown that this reaction requires the presence of factor XI. Factor XII has been purified by the method of Saito (Circ.Res.34,641,1974) , and factor XI has been purified by the method of Connellan, Castaldi and Muntz. (Haemostasis, in press 1977).Specific antibodies were developed in rabbits and the IgG from the antiserum was coupled to Sepharose. These factors could be extracted from plasma by the immobilized antibodies and could then be eluted by the use of alkali or guanidine HCL, thus resulting in a rapid purification of these factors.Platelets were washed free of loosely bound clotting factors and provided significant amounts of coagulant activity when ruptured by either freezing and thawing or by glycerol treatment. Platelets from a factor XI deficient patient also provided this activity. The platelet activity was extracted by passage through both Sepharose-heparin column and Sepharose-anti-factor XI column;the activity appeared to be identical to plasma factor XI.

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.

Sign in / Sign up

Export Citation Format

Share Document