Plasma Transglutaminase Research Articles

Safety and pharmacokinetics of recombinant factor XIII in healthy volunteers

Factor XIII (FXIII) is a plasma transglutaminase that covalently cross-links fibrin proteins to one another and to other proteins, increasing the mechanical strength of blood clots. Endogenous FXIII is the final enzyme in the clotting cascade and circulates as a heterotetramer comprising 2 FXIII-A subunits and 2 FXIII-B subunits. Recombinant human FXIII A2 (rFXIII) homodimer is produced in Saccharomyces cerevisiae. Upon injection, rFXIII combines with the free FXIIIB subunit that circulates in excess to form the rA2B2 tetramer. In this placebo-controlled, double-blind, multi-dose study, the safety, pharmacokinetics, and pharmacodynamics of rFXIII were studied in 24 healthy volunteers, who were randomized in 2 cohorts of 12 subjects each. In each cohort, 8 subjects received 5 daily intravenous doses of rFXIII (10 or 25 U/kg), and 4 subjects received placebo. Recombinant FXIII was well tolerated. No deaths or serious adverse events occurred. The type and frequency of adverse events showed no pattern or dose response. No clinically significant changes in haematology, serum chemistry, or urinalysis laboratory values were observed. No clinical coagulopathy or thrombosis was observed. Recombinant FXIII did not produce any anti-yeast or anti-FXIII antibodies. After 5 daily doses of rFXIII, accumulation indices indicated a 3 to 4fold accumulation of rFXIII in plasma. The elimination half-life, estimated for rFXIII as the heterotetramer, ranged from 228-346 hours for the 10U/kg dose group and 167-197 hours for the 25U/kg dose group. The safety, pharmacokinetic, and immunogenic profile of rFXIII suggests it may have potential use in patients with congenital or acquired FXIII deficiency.

Evaluation of Pro-Angiogenic Activity of Factor XIII (FXIII) in Ischemic Tissue, Heart Transplantation and FXIII-Deficient Mice.

FXIII, the last factor of the coagulation cascade, is a plasma transglutaminase that stabilizes fibrin clots by cross-linking fibrin chains. In addition to its role in coagulation, FXIII is essential for embryo implantation and wound healing - processes that involve angiogenesis. In our previous work (Dardik et al., Arterioscler. Thromb. Vasc. Biol. , 2003: 23: 1472) we showed that thrombin-activated FXIII (FXIIIa) exhibits pro-angiogenic activity that is associated with downregulation of thrombospondin (TSP-1). We next shoowed that the proangiogenic effect of FXIIIa is mediated by the phosphorylation and activation of VEGFR-2 receptor. Activated VEGFR-2 then initiates a signaling cascade leading to upregulation of cyclin D-1, Egr-1 and c-Jun, and downregulation of TSP-1 induced by c-Jun via WT-1 (submitted). In the current study we evaluated the pro-angiogenic effect of FXIIIa on cardiac ischemia (using rat myocardial ischemia model) or transplantation (usig murine neonatal cardiac allograft into pinnae ear transplant model). In addition, angiogenesis in FXIII-deficient mice was studied using in vivo Matrigel model. Following myocardial ischemia, a significant increase in new vessel formation was observed in the FXIIIa-injected as compared to saline-injected rats: 142±15/mm2 vs 64±15/mm2; respectively, p = <0.001. The increase in new vessel formation in the bFGF-treated animals (positive control) was similar to that observed in FXIIIa-treated mice (144±22/mm2; p = 0.6). Similarly, in the neonatal cardiac allograft transplant model the number of new vessels was higher in FXIIIa-treated animals than in controls 31.7±3.3 vs. 21.1±5.7; p<0.003. Moreover the function of the transplanted tissue as reflected by % heart beating area was significantly increased in FXIIIa-treated hearts than in the controls 64.7±14.9 vs. 40.3±21.2, respectively; p = 0.04.. In addition, the number of mice with >80% beating area was significantly higher in the FXIIIa-treated group that in the control: 46% vs 17%, respectively; p = 0.038. Histological analysis of the matrigel sections from FXIII-deficient mice showed significantly decreased number of new vessels compared to that of the control mice and the number of the vessels increased after treatment with FXIII (5.9±1.9 vr. 8.8±2.4 vr. 7.4±2.9, respectively, P=0.004). Similarly, the values of hemoglobin measured in the matrigel tissue were significantly higher in the control group as compared to that of the FXIII-deficient - group, increasding after treatment with FXIII (4.6 ± 2.5 mg/mg matrigel vs 1.3 ± 1.0 mg/mg matrigel vs. 4.4±0.6 mg/mg matrigel, respectively; p = 0.01. The results of this study confirm the pro-angiogenic activity of FXIIIa in ischemic tissues and clarify its role in tissue remodeling, thus, opening new therapeutic options for this compound with regard to clinical conditions where perfusion via development of new vessels is crucial.

Definition of the fine specificity of the monoclonal antibody 81D4: its reactivity with lysine and polyamine isopeptide cross-links

The 81D1C2 monoclonal antibody (Mab) directed against the N ɛ-(γ- l-glutamyl)- l-lysine isopeptide was found to cross-react on Enzyme Immuno Assay (EIA) with acylated lysines. Using a differential screening EIA procedure, a new Mab 81D4 was selected, which did not cross-react with acylated lysines but exhibited strong reactivity with N ɛ-(γ- l-glutamyl)- l-lysine formed by covalently coupling the γ-carboxyl of N αCBZ OtBu glutamic acid to Ɛ-NH 2 derivatized microtiter plates. When N ɛ-(γ- l-glutamyl)- l-lysine isopeptides were generated on γ-carboxyl derivatized plates, only lysine isopeptides with blocked α-amines were reactive, regardless of whether the bond formed by the amine blocking agent was a carbamate with carbobenzyloxychloride or an amide with acetic anhydride. The 81D4 Mab showed little or no affinity for free N ɛ-(γ- l-glutamyl)- l-lysine (IC 50>5 mM), for N 1 or N 4 mono(γ-Poly l-glutamyl)putrescine, and for N 1 mono(γ-Poly l-glutamyl)spermidine (IC 50>5 mM). However, when these same isopeptides were synthesized as cross-links between two protein chains— N ɛ-(γ- l-glutamyl)- l-lysine between Poly l-glutamate and Poly l-lysine; N 1 N 4- bis(γ-Poly l-glutamyl)putrescine, N 1 N 8- bis(γ-Poly l-glutamyl)spermidine between Poly- l-glutamate chains—very good reactivity was observed (IC 50 400 μM for lysine; 80 μM for putrescine and spermidine). In addition to the chemically synthesized isopeptide cross-links that were recognized by this Mab, the naturally occurring N ɛ-(ã- l-glutamyl)- l-lysine isopeptide cross-links in d-dimer, which are formed by the action of plasma transglutaminase (Factor XIII) on fibrin, were also detected on immunoblots using 81D4 as the primary antibody.

Factor XIIIa Incorporates Thymosin β4 Preferentially into the Fibrin(ogen) αC-Domains

It was shown recently that tissue transglutaminase and presumably plasma transglutaminase, factor XIIIa, can covalently incorporate into fibrin(ogen) a physiologically active peptide, thymosin beta(4) [(Huff et al. (2002) FASEB J. 16, 691-696]. To clarify the mechanism of this incorporation, we studied the interaction of thymosin beta(4) with fibrinogen, fibrin, and their recombinant fragments, the gamma-module (gamma-chain residues 148-411), and the alphaC-domain (Aalpha-chain residues 221-610) and its truncated variants by immunoblot and ELISA. No significant noncovalent interaction between them was detected in the absence of activated factor XIII, while in its presence thymosin beta(4) was effectively incorporated into fibrin and to a lesser extent into fibrinogen. The incorporation at physiological concentrations of fibrin(ogen) and factor XIII was significant with molar incorporation ratios of thymosin beta(4) to fibrinogen and fibrin of 0.2 and 0.4, respectively. Further experiments revealed that although activated factor XIII incorporates thymosin beta(4) into the isolated gamma-module and alphaC-domain, in fibrin the latter serves as the major incorporation site. This site was further localized to the COOH-terminal portion of the alphaC-domain including residues 392-610.

Structure and function of coagulogen, a clottable protein in horseshoe crabs.

Mammalian blood coagulation is based on the proteolytically induced polymerization of fibrinogens. Initially, fibrin monomers noncovalently interact with each other. The resulting homopolymers are further stabilized when the plasma transglutaminase (TGase) intermolecularly cross-links epsilon-(gamma-glutamyl)lysine bonds. In crustaceans, hemolymph coagulation depends on the TGase-mediated cross-linking of specific plasma-clotting proteins, but without the proteolytic cascade. In horseshoe crabs, the proteolytic coagulation cascade triggered by lipopolysaccharides and beta-1,3-glucans leads to the conversion of coagulogen into coagulin, resulting in noncovalent coagulin homopolymers through head-to-tail interaction. Horseshoe crab TGase, however, does not cross-link coagulins intermolecularly. Recently, we found that coagulins are cross-linked on hemocyte cell surface proteins called proxins. This indicates that a cross-linking reaction at the final stage of hemolymph coagulation is an important innate immune system of horseshoe crabs.

Effect of porcine plasma protein and setting on gel properties of surimi produced from fish caught in Thailand

Effects of porcine plasma protein (PPP) and high temperature setting on gel properties of surimi from bigeye snapper, bigeye croaker, threadfin bream and barracuda were investigated. PPP was effective in increasing breaking force and deformation of kamaboko gels set at 40°C for 30 min and heated at 90°C for 20 min. The optimum levels of PPP were 0.5, 0.5, 1.5 and 1.5 g/100 g and the optimum setting times were 2, 1.5, 1.5 and 2 h for bigeye snapper, bigeye croaker, threadfin bream and barracuda surimi, respectively. However, the addition of PPP significantly decreased whiteness ( P<0.05). An increase in gel-forming ability of surimi with PPP coincided with a decrease in solubility in mixture of SDS, urea and β-mercaptoethanol, indicating the formation of nondisulfide covalent bond induced by both endogenous and plasma transglutaminase. The results supported that PPP improve the gelation of surimi in combination with setting.

Novel proangiogenic effect of factor XIII associated with suppression of thrombospondin 1 expression.

Factor XIII (FXIII), a plasma transglutaminase that stabilizes fibrin clots at the final stages of blood coagulation by crosslinking fibrin monomers, is essential for embryo implantation and participates in tissue remodeling and wound healing, processes that involve angiogenesis. The aim of our study was to analyze the effect of FXIII on angiogenesis using in vitro and in vivo models and to examine the role of FXIII in the basic steps of angiogenesis, ie, migration, proliferation, and apoptosis/cell survival. In the Matrigel tube formation model, only FXIIIa caused a dose-dependent enhancement of array formation. This proangiogenic effect was not associated with alterations in vascular endothelial growth factor (VEGF) protein levels nor VEGF or VEGFR2 mRNA levels. FXIIIa, but not nonactivated or transglutaminase-inactivated FXIII, significantly enhanced endothelial cell migration and proliferation and inhibited apoptosis. After treatment of HUVECs with FXIIIa, almost complete disappearance of mRNA of thrombospondin 1 (TSP-1) and a marked reduction in the secretion of TSP-1 protein were observed. A reduction in TSP-1 protein synthesis, although to a lesser extent, was observed on treatment of microvascular endothelial cells with FXIIIa. In a rabbit cornea model, injection of FXIIIa caused neovascularization associated with almost complete disappearance of TSP-1 in the cornea. These results show that FXIIIa exhibits a novel proangiogenic activity that is associated with downregulation of TSP-1 and also involves stimulation of endothelial cell proliferation and migration and inhibition of apoptosis. These findings might shed light on the mechanism by which FXIII mediates tissue repair and remodeling.

Transglutaminase Inhibitor from Milk

ABSTRACT Cross‐linking experiments of skimmed bovine milk with bacterial transglutaminase isolated from Streptoverticillium mobaraense showed only some degree of formation of high‐molecular‐weight casein polymers. Studies on the nature of this phenomenon revealed that bovine milk contains an inhibitor of transglutaminase activity. Removal of the casein and whey proteins from the milk resulted in a protein‐poor fraction that still inhibited transglutaminase activity at cross‐linking of β‐casein and in several activity assays of transglutaminase. The inhibitor was partially purified by column chromatography and appeared to be a heat labile low molecular weight component. Inhibition of transglutaminase activity was observed with microbial transglutaminase, plasma transglutaminase and guinea pig liver transglutaminase. The inhibiting activity was found in bovine, goat, sheep, and human milk, but could not be detected in horse milk.

Type XVI Collagen is Expressed in Factor XIIIa+ Monocyte-Derived Dermal Dendrocytes and Constitutes a Potential Substrate for Factor XIIIa

We have previously reported that connective tissue cells in the superficial dermis preferentially express alpha1(XVI) collagen rather than those in the lower dermis. Double immunofluorescence labeling using the antibodies for alpha1(XVI) collagen and factor XIIIa (plasma transglutaminase), which is a marker of dermal dendrocytes, demonstrated that both antibodies reacted with the same cells in the superficial dermis of normal skin as well as the lesional skins of dermal dendrocyte-related disorders, dermatofibroma, and psoriasis. Dermal dendrocytes are considered to be established by a culture of peripheral blood monocytes in the presence of granulocyte macrophage-colony stimulating factor and interleukin-4. Reverse transcription--polymerase chain reaction, metabolic labeling, and immunofluorescence studies demonstrated that treatment of CD14+ peripheral blood monocytes with granulocyte macrophage-colony stimulating factor/interleukin-4 over a period of 8 d resulted in the induction of alpha1(XVI) collagen as well as factor XIIIa. The physiologic significance of colocalization of alpha1(XVI) collagen and factor XIIIa in the tissue and their coordinate induction in CD14+ monocyte-derived dendritic cells in vitro was studied. Considerable incorporation of [3H]putrescine by factor XIIIa into recombinant noncollagenous domain (NC) 11 but not into collagenous domain (COL) 1.NC1 domain of the alpha1(XVI) polypeptide was found. Incubation of recombinant NC11 of alpha1(XVI) polypeptide with factor XIIIa in vitro produced a covalent cross-linking complex on sodium dodecylsulfate-polyacrylamide gel electrophoresis. The results indicate that alpha1(XVI) collagen is constitutively expressed by most dermal dendrocytes in the skin and dendritic cells differentiated from peripheral blood monocytes in vitro. Type XVI collagen is expressed in factor XIIIa+ dermal dendrocytes and may form an intermolecular cross-linking through NC11 domain by the reaction catalyzed by factor XIIIa contributing to the structural integrity of factor XIIIa+ dendritic cell-rich tissues.

Evaluation of Transglutaminase from Pig Plasma on the Quality of Milk Curd

The purpose of this study was to examine the effect of different levels of crude pig plasma transglutaminase (TGase) on the quality of milk curd. Different levels (0, 0.1, 0.2, 0.4, 0.8 and 1.0%) of crude pig plasma TGase was added to fresh milk, individually, and then incubated at 35°C. The time of milk curdling was recorded. Simultaneously, rheological properties, L value and microstructure of the samples were measured and observed. The results showed that the time of milk curdling decreased and the curd strength of milk curd increased with increasing of TGase (p<0.05). The softness of milk curd had the highest value when 0.8% TGase was added (p<0.05). However, L value of milk curds was not significantly different among all treatments. The microstructure of milk curd observed by scanning electron microscopy (SEM) became more dense with TGase level increased. (Asian-Aust. J. Anim. Sci. 2002. Vol 15, No. 1 : 106-110)

The Biological Functions of MBL-Associated Serine Proteases (MASPs)

The Mannose-binding lectin-associated serine proteases (MASPs) have been the subject of intensive research particularly over the past 10 years. First one, then two, and currently 3 MASPs have been characterized. Initially it was thought likely that the MBL + MASPs system would resemble very closely the C1 complex of the complement classical pathway, and that MASP1 and MASP2 would have similar activities to their classical pathway homologues C1r and C1s. MASP2 does certainly have similar activities to C1s, but MASP1 does not have the activities of either C1r or C1s. MASP1 has been thought to act on the complement system by cleaving C3 directly, but work with recombinant and purified native MASP1 shows that direct C3 cleavage by this protease is very slow, and may not be biologically significant. MASP1 and MASP2 appear not to have such a narrow specificity as C1r and C1s, and may have significant substrates other than complement proteins. As an example, MASP1 does cleave fibrinogen, releasing fibrinopeptide B (a chemotactic factor) and also cleaves and activates plasma transglutaminase (Factor XIII). These reactions are also relevant to defence against microorganisms, and may represent a biologically significant action of MASP1.

Purification of transglutaminase and its effects on myosin heavy chain and actin of spent hens

The purpose of this study was to purify pig plasma transglutaminase (TGase) and examine its effects on the myosin heavy chain and actin of the breast muscles from spent hens at different temperatures. TGase (0.3 units/mg) was added to myofibrillar proteins solution (0.5 ml) at 4°C for 0, 4, 8, 12, 24 and 48 h; at 25°C for 0, 1, 2, 4, 8, 12, 24 and 48 h; at 37°C for 0, 5, 10, 30 and 60 min. The results of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE) showed that plasma TGase was composed of units of molecular weights approximately 75,000 and 80,000. TGase added to the myofibrillar proteins solution indicated that the concentration of the myosin heavy chain and actin decreased when incubated at 4°C for 48 h and when incubated at 25°C for 2 h. Moreover, the relative intensity determined by scanning densitometry of the SDS–PAGE gel indicated that the myosin heavy chain and actin concentration decreased to 45 and 64%, respectively. In addition, the relative intensity of the myosin heavy chain and actin declined to 7 and 63%, respectively, when incubated at 37°C for 5 min. The relative intensity of both the myosin heavy chain and actin decreased with time when incubated at 25 and 37°C.

PORCINE PLASMA PROTEINS AS GEL ENHANCER IN BIGEYE SNAPPER (PRIACANTHUS TAYENUS) SURIMI

Cohn 's fraction I-Sfiom porcine plasma showed the highest transglutaminase activity, compared to fractions I, II+III, IV, IV-I. The optimum temperature for incorporating monodancylcadaverine into dimethylated casein was 45C. Plasma transgfutaminase in fraction I-S was activated by calcium chloride but was inhibited by N-ethylmaleimide, ethylenediaminetetraacetic acid, and ammonium chloride. The addition of fraction I-S into bigeye snapper surimi resulted in a substantial increase in gel breaking force and deformation, particularly in the presence of calcium chloride and thrombin. No changes in whiteness and water holding capacity were observed in surimi gel with the addition of 0-0.5% of fraction IS. Fraction I-S was found to catalyze nondisulJide covalent cross-linking of myosin heavy chain. The combination of endogenous and plasma transglutaminase enhanced surimi gelation.

Purification and substrate specificity of transglutaminases from blood and Streptoverticillium mobaraense.

A procedure for a fast and simple purification of bovine plasma transglutaminase was developed, which resulted in a homogeneous enzyme preparation. Two different procedures were developed for the purification of pig erythrocyte transglutaminase, both of which resulted in partial purification. Both enzymes were used in cross-linking reactions of alpha-lactalbumin, beta-lactoglobulin, bovine serum albumin, casein, hemoglobin, glycinin, and myosin. The substrate specificity was compared to that of bacterial transglutaminase isolated from Streptoverticillium mobaraense. The bacterial transglutaminase caused cross-linking of a wider range of proteins and, thus, exhibited a lower substrate specificity than the blood transglutaminases. In addition, differences exist in the necessity of the addition of reducing agents. These differences allow specific applications of blood and bacterial transglutaminases at protein cross-linking in single or complex protein systems.

The role of plasma transglutaminase (F XIII) in wound healing of complicated pressure sores after spinal cord injury

A case report. To demonstrate stimulating action of F XIII in wound healing of complicated pressure sores. A Spinal Cord Injury Center in Germany. Clinical exam, clinical and photographic wound control, biochemical serum monitoring. Recurrent pressure sores in plegic patients are common complications requiring long-standing conservative or operative therapy. Additional risk factors such as diabetes increase the complication rate for surgery. Surgery itself may be difficult in recurrent pressure sores due to limited remaining soft tissues. We report the case and treatment of a 47-year-old patient with long-standing and recurrent ulcers and complications after flap surgery. As a final option we added plasma transglutaminase (factor XIII) to our treatment scheme which changed the course of the disease dramatically and we achieved complete and rapid healing. Our experience suggests that F XIII has a positive role in treating pressure sores as shown already in several other surgical fields. Its use is giving the surgeon an additional tool in complicated cases.

Physiopathology and Regulation of Factor XIII

Factor XIII is a plasma transglutaminase. Transglutaminases are at least 8 enzymes which cross-link a number of proteins. This type of reaction not only enhances the original functions of substrate proteins, but also adds new functions to them. Factor XIII in plasma is a tetramer (A2B2), and the A subunit contains the active site. Although transglutaminases are homologous, the nucleotide sequences in their 5'-flanking region differ significantly. Accordingly, transcription factors play a major role in the cell type-specific expression of each transglutaminase. A variety of missense and nonsense mutations, and deletions/insertions with or without out-of-frame shift/premature termination and splicing abnormalities have been identified in the genes for A and B subunits in factor XIII deficiency. In some cases, the mRNA level of the A or B subunit was severely reduced. Molecular and cellular bases have also been explored by expression experiments and by molecular modeling. In most cases, impaired folding and/or conformational change of the mutant A or B subunit leads to both intra- and extra-cellular instability, which is responsible for factor XIII deficiency.

Factor XIII Improves Gastric Stress Lesions in Rats

Background/Aims: Tissue transglutaminase has been reported to be involved in the healing of experimental gastric ulcer; nevertheless, other type(s) of transglutaminase could be involved. The present experiments aimed at examining whether plasma transglutaminase (factor XIIIa) contributes to such healing and at evaluating whether factor XIII supplementation improves gastric mucosal lesions. Methods: The healing effect of 200 U/kg of factor XIII administered intravenously was examined using a water immersion restraint rat model of stress gastric damage. The rats were sacrified 0, 2, 4, and 12 h after stress. The gastric mucosa was examined macroscopically and microscopically, and the transglutaminase activities were assayed in serum and gastric mucosa. Factor XIIIa and tissue transglutaminase protein levels in the gastric mucosa were analyzed by immunoblot. Immunohistochemistry was used to identify the location of tissue transglutaminase, factor XIIIa, and fibronectin in the gastric mucosa. Results: The transglutaminase activity, reduced by stress in the gastric mucosa, increased up to 12 h after stress, peaking at 4 h, when the ulcer index significantly decreased. The serum transglutaminase level was low at all time points. Exogenous administration of factor XIII allowed a faster reduction of the ulcer index that was coincident with an increased transglutaminase activity in the mucosa. Both tissue transglutaminase and factor XIIIa protein levels were reduced by 6 h of stress and increased after factor XIII administration. Immunohistochemistry showed a colocalization of both factor XIIIa and tissue transglutaminase with fibronectin in the extracellular matrix of the damaged area. Conclusions: Two forms of transglutaminase are involved in the healing of stress-induced gastric erosions, and factor XIII administration allows faster gastric mucosa healing.

Cross-linking of Wild-type and Mutant α2-Antiplasmins to Fibrin by Activated Factor XIII and by a Tissue Transglutaminase

Human alpha(2)-antiplasmin (alpha(2)AP), the main inhibitor of plasmin-mediated fibrinolysis, is a substrate for plasma transglutaminase, also termed activated factor XIII (FXIIIa). Of 452 amino acids in alpha(2)AP, only Gln(2) is believed to be a fibrin-cross-linking (or FXIIIa-reactive) site. Kinetic efficiencies (k(cat)/K(m)((app))) of FXIIIa and the guinea pig liver tissue transglutaminase (tTG) and reactivities of Gln substrate sites were compared for recombinant wild-type alpha(2)AP (WT-alpha(2)AP) and Q2A mutant alpha(2)AP (Q2A-alpha(2)AP). [(14)C]Methylamine incorporation showed the k(cat)/K(m)((app)) of FXIIIa to be 3-fold greater than that of tTG for WT-alpha(2)AP. With FXIIIa or tTG catalysis, [(14)C]methylamine was incorporated into Q2A-alpha(2)AP, indicating that WT-alpha(2)AP has more than one Gln cross-linking site. To identify transglutaminase-reactive sites in WT-alpha(2)AP or Q2A-alpha(2)AP, each was labeled with 5-(biotinamido)pentylamine by FXIIIa or tTG catalysis. After each labeled alpha(2)AP was digested by trypsin, sequence and mass analyses of each labeled peptide showed that 4 of 35 Gln residues were labeled with the following reactivities: Gln(2) > Gln(21) > Gln(419) > Gln(447). Q(2)A-alpha(2)AP was also labeled at Gln(21) > Gln(419) > Gln(447), but became cross-linked to fibrin by FXIIIa or tTG at approximately one-tenth the rate for WT-alpha(2)AP. These results show that alpha(2)AP is a better substrate for FXIIIa than for this particular tTG, but that either enzyme involves the same Gln substrate sites in alpha(2)AP and yields the same order of reactivities.

Evaluation of transglutaminase on the quality of low-salt chicken meat-balls

The purpose of this study was to examine the effect of different concentrations of crude pig plasma transglutaminase (TGase) at 0, 0.05, 0.1, 0.2, 0.4 and 1.0% on the quality of low-salt chicken meat-balls. Yield, color, gel strength, microstructure and sensory evaluation were measured. The results showed that both the yield and gel strength of the meat-balls increased as TGase supplement increased ( P<0.05) without any obvious effect on color. Scanning electron microscopy (SEM) indicated firmer and more regular gel structures with increasing TGase addition. Texture, juiciness and overall acceptability as judged by sensory evaluation were not statistically affected until the level of TGase reached 1.0% ( P<0.05).

Racial and genetic determinants of plasma factor XIII activity.

Factor XIII (F XIII), a plasma transglutaminase, is essential for normal hemostasis and fibrinolysis. Plasma F XIII consists of two catalytic A (F XIIIA) and two non-catalytic B (F XIIIB) subunits. Activated F XIII is involved in the formation of fibrin gel by covalently crosslinking fibrin monomers. As the characteristics of the fibrin gel structure have been shown to be associated with the risk of coronary heart disease (CHD), F XIII activity may play a seminal role in its etiology. In this investigation, we determined plasma F XIII activity in two racial groups, including Asian Indians (n = 258) and Chinese (n = 385). Adjusted plasma F XIII activity was significantly higher in Indian men (142 vs. 110%; P<0.0001) and women (158 vs. 111%; P<0.0001) than their Chinese counterparts. As compared to Indians where the distribution of F XIII activity was almost normal, in Chinese it was skewed towards low activity. In both racial groups, bivariate and multivariate analyses showed strong correlation of F XIII activity with plasma fibrinogen and plasminogen levels. Race explained about 25% of the variation in F XIII activity even after the adjustment of significant correlates. We also determined the contribution of common genetic polymorphisms in the F XIIIA and F XIIIB genes in affecting plasma F XIII activity. Both loci showed significant and independent effects on plasma F XIII activity in Indians (F XIIIA, P< 0.01; F XIIIB, P<0.05) and Chinese (F XIIIA, P<0.0001; F XIIIB, P<0.13) in a gene dosage fashion. This study shows that both racial and genetic components play a significant role in determining plasma F XIII activity, and consequently it may affect the quantitative risk of CHD.