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At least 19 recordsLinked to original sources

PubMed1975

Cross-linking of fibrinogen and fibrin by fibrin-stablizing factor (factor XIIIa).

Factor XIIIa catalyzed intermolecular cross-linking of fibrinogen at initial rates that varied in direct (first order) proportion to the fibrinogen concentration, which differed from the well known zero order relationship in fibrin cross-linking. Preferential cross-linking of gamma-chains occurred with both substrates. The differences in rates and order of reaction were attributed mainly to effect of self-alignment of the gamma-chains in fibrin which enabled the cross-linking enzyme to interact with paired chains as a single rather than two independent entities. Studies on mixtures of fibrinogen and fibrin indicated factor XIIIa had near equal affinities for the two substrates. At low concentrations with which cross-linking of fibrinogen proceeded sluggishly compared to fibrin, fibrinogen inhibited stabilization of fibrin clots by competitively partitioning factor XIIIa away from the fribin. Additional inhibition arose from cross-linking of fibrin in soluble combination with fibrinogen in mixtures containing fibrinogen in large excess over fibrin. The observations demonstrate ways in which fibrinogen normally helps to suppress both polymerization and cross-linking of small amounts of fibrin produced within the circulation. At very high concentrations above 30 mg. per milliliter, fibrinogen underwent cross-linking at faster initial rates than the cross-linking of fibrin. Rapid cross-linking of concentrated fibrogen raises the possibility that filtration enrichment may be a factor contributing to abnormal formation of the highly insoluble fibrinogen deposits occurring in atheromatous tissue.

Animals↗

PubMed1988

Fibrous papule: a tumor of fibrohistiocytic cells that contain factor XIIIa.

Factor XIIIa, a blood coagulation factor, has been found in a variety of cell types, including dendritic reticulum cells and fibroblast-like mesenchymal cells. We hypothesized that fibrous papule, a lesion of uncertain histogenesis, was composed of dermal stellate cells and in this report demonstrate that this neoplasm consists of cells that contain this factor. Nevus cells do not contain factor XIIIa.

Biopsy↗

PubMed2002

[Inhibitors of factor XIIIa].

Factor XIIIa (FXIIIa) catalyzes the covalent crosslinking of fibrin polymers and incorporation of proteins into the fibrin network and thus confers on the thrombus additional structural stability and relative resistance to plasmin-mediated degradation. Moreover, FXIIIa is involved in other physiological and pathophysiological processes such as wound healing and arteriosclerosis. Selective FXIIIa inhibitors may be a valuable tool for evaluation of the various functions of FXIIIa and their pharmacological control. This paper presents an overview of the inhibitors of FXIIIa. Analogues of natural FXIIIa substrates - including glutamine containing peptides and low molecular weight substituted alkylamines - are incorporated into the fibrin network and thus prevent crosslinking of fibrin. Naturally occurring, direct inhibitors of FXIIIa have been isolated from a leech species and microorganisms. With effective concentrations in the nanomolar range the peptide tridegin is the most potent FXIIIa inhibitor up to now. The majority of the synthetic, low molecular Weight inhibitors bind covalently to Cys314 at the active site of FXIIIa. Besides the relatively nonspecific thiol reagents, azol derivatives, azolium salts and related substances are described as specific inhibitors of FXIIIa. They inhibit the activity of FXIIIa at nanomolar concentrations. Animal experiments have demonstrated improved thrombolysis by a plasminogen activator in combination with a FXIIIa inhibitor.

Arteriosclerosis↗

PubMed1991

[Immunohistochemical study of the distribution of collagens (type I, III, IV), fibronectin, factor XIIIa and factor VIII related antigen in alcoholic liver disease].

Histopathological changes in alcoholic liver disease are characterized by Mallory body, fatty liver, and peculiar fibrosis such as pericentral fibrosis, pericellular fibrosis, stellate fibrosis originating from Glisson's sheath, and swelling of the liver cells. However, the mechanism of fibrosis remains unknown. The author tried to demonstrate, by the immunohistochemical method, fibronectin (FN), collagen type I, III, IV (IC, IIIC, IVC), factor XIIIa, and factor VIII related antigen (VIII RAg), all of which are related to the process of wound healing, in order to clarify the mechanism of fibrosis in alcoholic liver disease. With the progress of fibrosis, FN was demonstrated in the sinusoidal wall, as well as the portal areas and around the central veins. IC and IIIC were positive in the fibrotic area extending into the lobules and pericellular fibrosis. IVC was similar to FN and was positive in the sinusoidal wall. Scattered XIIIa positivity was recognized in the fibrotic areas, especially in the foci of active fibrosis. VIII RAg was positive in the capillary vessels invading the lobules through the limiting plates. The distribution of these proteins in the fibrotic area in alcoholic liver disease closely resembled that in the wound healing process. It is concluded that fibrosis occurring in the periportal area is essentially the same as in the wound healing process and the mechanism of fibrosis in alcoholic liver disease is thought to be active fibroplasia.

Adult↗

PubMed1996

Immunohistochemical detection of factor XIIIa and factor XIIIs in synovial membranes of patients with rheumatoid arthritis or osteoarthritis.

In spite of differences in etiology, RA and OA lead to astonishingly similar synovitic alterations. Fibroblastic transformation of the synovial membrane and an increase in monocytes constitute a rare but highly characteristic feature of RA. Monocytes synthesize factor (F) XIII, implying that FXIII (a and s) in synovial tissue might help to differentiate between RA and OA. Biopsies were obtained at open surgery from 98 unselected patients with the clinical diagnosis of RA (n = 54) or OA (n = 44). In a three-stage (ABC) immunoperoxidase technique, polyclonal antisera against factor XIIIa and factor XIIIs were investigated. Compared to OA sections. RA synovium showed more FXIIIa-positive cells-monocytes, fibrocytes, fibroblasts and synovial lining cells. In the subsynovial layer, band-like structure of FXIIIa-stained cells was observed in 27.8% of the RA patients, but in only one OA specimen. Higher proportions of FXIIIa-positive monocytes, macrophages, histiocytes and fibroblasts, as well as positive Langhans giant cells and vascular wall regions (except endothelial cells), were observed in RA. OA specimens revealed more intense FXIIIa labeling of these cells with a lower percentage of stained cells. Overall, labeling with FXIIIs antibody resulted in less intense staining. In conclusion, distinction between synovitis caused by RA and synovitis due to OA is possible, as the former show higher numbers of FXIIIa-positive cells, including monocytes, fibroblasts, fibrocytes and synovial lining cells. Further more, RA tissue is stained less intensely than OA tissue. There is evidence for continuous excretion of FXIII in the synovial membrane by the above-mentioned cell systems.

Adolescent↗

PubMed2003

Staphylococcus aureus fibronectin-binding protein serves as a substrate for coagulation factor XIIIa: evidence for factor XIIIa-catalyzed covalent cross-linking to fibronectin and fibrin.

In this study, we have investigated the interactions of a Staphylococcal recombinant fibronectin-binding protein A (rFnbA) with fibronectin, fibrinogen, and fibrin. Using analytical size-exclusion chromatography, we evaluated the stoichiometry of reversible binding of FnbA to fibronectin and demonstrated that, in solution, it can accommodate at least two molecules of fibronectin. Results of ELISA experiments demonstrated that rFnbA binds with equally high affinity to both immobilized fibrinogen and fibrin. When included into a thrombin-induced fibrin polymerization reaction, rFnbA strongly inhibited fibrin assembly in a dose-dependent manner. In this study, we have shown that rFnbA can act as a substrate for coagulation factor XIIIa. Factor XIIIa catalyzes the incorporation of amine donor (dansylacadaverine) and amine acceptor (peptide patterned on the N-terminal sequence of fibronectin) synthetic probes into rFnbA, suggesting that it serves as a bifunctional substrate containing reactive glutamine and lysine residues. We have demonstrated that the reversible complex formed by rFnbA and fibronectin or rFnbA and fibrin is covalently stabilized by the transglutaminase action of factor XIIIa. Incubation of rFnbA in the presence of either of its ligands and factor XIIIa results in the introduction of intermolecular epsilon-(gamma-glutamyl)lysine isopeptide bond(s) and the formation of high molecular mass heteropolymers. These findings suggest a novel mechanism by which pathogenic Staphylococcus aureus may utilize the transglutaminase activity of factor XIIIa for attachment to soluble proteins, cell surfaces, and matrixes.

Adhesins, Bacterial↗

PubMed2002

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.

Adult↗

PubMed1997

An unusual soft tissue tumor with features of angiomatoid malignant fibrous histiocytoma composed of bimodal CD34 and factor XIIIa positive dendritic cell subsets. CD34 and factor XIIIa in angiomatoid MFH.

CD34 and factor XIIIa (FXIIIa) antibodies delineate subsets of embryonic dendritic stromal stem cells that persist in adult mesenchyme. CD34 stains interstitial and adventitial dendritic cells that may function as multipotential precursor cells. FXIIIa+ dendrophages are tissue histiocytes active in tissue repair. Angiomatoid malignant fibrous histiocytoma (AMFH) is an enigmatic fibrohistiocytic tumor with limited vascular features. We examined an unusual soft tissue tumor from the nasolabial subcutis of a 57 year old man that showed histologic features of AMFH. Most of the tumor cells expressed CD34. 10-30% of the tumor cells were FXIIIa+ dendrophages. Sinusoidal areas were largely composed of FXIIIa+ cells that also expressed HLA-DR and CD68 suggesting macrophage differentiation. CD31 and Factor VIII antigen highlighted capillaries and single cells among the CD34+ tumor cells. The vessels had actin+ myopericytes and there were single actin+ tumor cells. Electron microscopy showed primitive dendritic cells and fewer histiocyte-like cells. The Ki 67 index was 15% including both FXIIIa+ and CD34+ cells. The patient is disease free three years after wide excision. We conclude that this AMFH-like neoplasm is a fibrohistiocytic tumor in which CD34+ fibroblast-like precursors and FXIIIa+ tissue dendrophages combine to build both sinusoidal tissue with endothelial and macrophage elements as well as capillary vascular tissue that is invested with myopericytes. Study of additional AMFH lesions from this standpoint is desirable.

Antigens, CD34↗

PubMed1980

Structural features of glutamine substrates for human plasma factor XIIIa (activated blood coagulation factor XIII).

The action of human plasma factor XIIIa (thrombin-activated blood coagulation factor XIII) and guinea pig liver transglutaminase on purified caseins, fibrin, the derivatized gamma chain of fibrin, and a number of synthetic glutamine peptides, and peptide derivatives is reported. There are wide variations in the properties of the individual proteins and peptides as substrates for amine incorporation by the two transglutaminases. beta-Casein and several of its derivatives are excellent substrates for factor XIIIa. However, beta-casein is a relatively poor substrate for the liver enzyme. The primary site of amine incorporation by factor XIIIa in beta-casein was identified as glutamine 167. This was accomplished by labeling with fluorescent amine followed by proteolytic digestion and identification of labeled peptides. An 11-residue peptide and a 15-residue peptide, each containing 1 glutamine residue and each modeled after the primary site of amine incorporation in beta-casein, were prepared. A 13-residue peptide modeled after the primary crosslinking site in fibrin gamma chain was also prepared. Each of these polypeptides proved to be an efficient substrate for factor XIIIa and displayed significantly better substrate properties than a number of small glutamine peptide derivatives that are good substrates for liver transglutaminase.

Amino Acid Sequence↗

PubMed1986

Factor V is a substrate for the transamidase factor XIIIa.

Coagulation Factor V (Mr = 330,000), upon cleavage by thrombin, produces Factor Va, which is composed of two subunits with Mr values of 94,000 and 74,000, along with two activation fragments possessing no known function. Studies were undertaken to assess the ability of the transamidase Factor XIIIa to covalently incorporate the lysine analogs [3H]putrescine and dansylcadaverine into the thrombin-cleaved (activated) and unactivated forms of human and bovine Factor V. The incorporation of either probe into thrombin-activated Factor V proceeded at an initial rate approximately twice that for unactivated Factor V. The extent of the incorporation of [3H]putrescine or dansylcadaverine into activated or unactivated human Factor V was identical; 4 mol of either probe per mol of Factor V. In the case of bovine Factor V, however, while 4 mol of probe were bound per mol of the unactivated pro-cofactor, 5 mol of either lysine analog were covalently linked to 1 mol of thrombin-cleaved Factor V. Polyacrylamide gel fluorography, immunoaffinity chromatography, and immunoprecipitation identified the largest activation fragment of human Factor V (Mr = 150,000) and bovine Factor V (Mr = 120,000) to contain the sites of incorporation of the covalently bound probes. High molecular weight, apparently covalent polymers of Factor V were produced by the action of Factor XIIIa on activated and unactivated human or bovine Factor V. The absence of either probe in the reaction mixtures did not appear to allow an enhancement of protein polymerization.

Animals↗

PubMed1996

Myxoid dermatofibrohistiocytoma: an indolent post-traumatic tumor composed of CD34+ epithelioid and dendritic cells and factor XIIIa+ dendrophages.

Factor XIIIa+ dendrophages and CD34+ "deep dermal dendrocytes" are distinct subsets of embryonic dermal dendritic stem cells that persist in interstitial and adventitial sites in adult dermis. We encountered a unique myxoid dermal tumor composed of these two cell types. It arose after trauma to the thumb of a 49-year-old man and was locally excised. The patient is without recurrence at 18 months. The disc-shaped tumor was lobulated, yellow, and mucoid and involved the margins. A fibrillar myxoid stroma contained mast cells, wispy collagen with medium-to-small vessels, and loosely deployed small eosinophilic tumor cells. The tumor cells were amitotic and had oval or bean-shaped, bland nuclei; some cells were binucleated. The cells were epithelioid or dendritic with bipolar, stellate, and racquet-shaped cytosomes whose tapering cell processes blended with fibrillar collagen. Vacuolated epithelioid cells focally formed vessel-like luminal structure. All cells strongly expressed vimentin. Thirty percent of the tumor cells were elongated, dendritic factor XIIIa+ cells whose dendritic processes enshrouded mast cells or FXIIIa-negative tumor cells. A subset of the FXIIIa+ cells also expressed MAC387 and lysozyme. The other 70% of the cells were CD34+. Many CD34+ cells were epithelioid with strong membrane and vacuolar decoration. Some CD34+ epithelioid cells had globular cytoplasmic inclusions. Other CD34+ cells were dendritic with multipolar fibrobroblast-like cytosomes and weaker CD34+ membrane decoration. Actin and S-100 were negative. Ki 67 was expressed in 1% of the tumor cells. Double stains for CD34 and Ki 67 showed that both CD34+ cells and FXIIIa+ dendrophages were Ki 67+, as were many papillary dermal vessel endothelial cells. The composition of the tumor by mast cells, FXIIIa+ dendrophages, and CD34+ primitive cells recapitulates the dermal microvascular unit. We propose the descriptive term myxoid dermatofibrohistiocytoma (MD) for this novel tumor. It appears to be an unusual response by dermal dendritic cells, perhaps due to continued stimulation by post-traumatic cytokines. Clarification of its biology and nosology awaits identification and study of more cases.

Antigens, CD34↗

PubMed1978

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

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.

Blood Platelets↗

PubMed1993

The tissue distribution of factor XIIIa positive cells.

Factor XIIIa has previously been used to identify a subgroup of dermal dendritic cells which co-express class II MHC antigens, intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function associated antigen-1 (LFA-1), leading to the suggestion that they play a central role in skin immune responses. To examine whether these cells may also be important constituents of other organs we have performed an immunohistochemical survey of normal tissue to identify the distribution of this factor XIIIa positive subgroup of dendritic cells in humans. Routine tissue sections from a range of organs were assayed for factor XIIIa expression using a standard peroxidase-antiperoxidase (PAP) immunolabelling technique. Dendritic, factor XIIIa positive cells were observed in all tissues studied, but were most numerous in skin and mucosal tissues (gastrointestinal tract, bladder). They were also observed associated with epithelial structures in lung and kidney, but were only rarely observed in liver, thyroid, testis and spleen. The distinctive distribution of these cells is consistent with an important role for them in immune responses at those sites.

Connective Tissue↗

PubMed1998

Identification of a large deletion, spanning exons 4 to 11 of the human factor XIIIA gene, in a factor XIII-deficient family.

Inherited deficiency of factor XIIIA subunit (FXIIIA) is an autosomal recessive disorder that is characterized by a life-long bleeding tendency and complications in wound healing. Molecular genetic studies have shown the deficiency can be due to small sequence changes within the FXIIIA gene, such as point mutations or microdeletions. On molecular analysis of the FXIIIA gene in an FXIII-deficient patient, of United Kingdom origin, we identified a putative homozygous missense mutation, Arg408Gln. However, the father of this patient is homozygous normal for arginine at codon 408. Having proved paternity in this pedigree by microsatellite analysis, we examined the FXIIIA RNA of the patient by reverse transcriptase-polymerase chain reaction and found the paternal allele to lack exons 4 through 11 inclusive. Hence, a huge deletion extending from intron 3 to intron 11 and the Arg408Gln mutation are jointly responsible for FXIIIA deficiency in this family. This is the first finding of such a large deletion in the FXIIIA gene.

Adult↗

PubMed2001

Interleukin-1 induces pro-mineralizing activity of cartilage tissue transglutaminase and factor XIIIa.

Two transglutaminases (TGases), Factor XIIIa and tissue TGase (tTGase), are expressed in temporal-spatial association with matrix calcification in growth plates. Meniscal and articular cartilage matrix calcification are prevalent in osteoarthritis (OA) and aging. Here, we demonstrated up-regulation of tTGase and Factor XIIIa in superficial and deep zones of knee OA articular cartilage and the central (chondrocytic) zone of OA menisci. Transforming growth factor-beta and interleukin (IL)-1beta induced Factor XIIIa and tTGase expression in cartilage and meniscal organ cultures. Thus, we studied TGase activity. Donor age-dependent, OA severity-related, and IL-1-induced increases in TGase activity were demonstrated in both knee menisci and cultured meniscal cells. Meniscal cell TGase activity was stimulated by nitric oxide donors and tumor necrosis factor-alpha, but transforming growth factor-beta did not stimulate TGase activity. The iNOS inhibitor N-monomethylarginine (NMMA) and an inhibitor of tumor necrosis factor receptor-associated factor (TRAF)2 and TRAF6 signaling (the zinc finger protein A20) suppressed IL-1 induction of TGase activity. Increased Factor XIIIa and tTGase activities, achieved via direct transfection of chondrocytic TC28 and meniscal cells, both induced matrix apatite deposition. Thus, Factor XIIIa and tTGase activities were increased in aging, degenerative cartilages and induced by IL-1. Because TGase activity promoted apatite deposition, our findings potentially implicate inflammation in the pathogenesis of cartilage matrix calcification.

Cadaver↗

PubMed1993

Cultured Kaposi's sarcoma cell lines express factor XIIIa, CD14, and VCAM-1, but not factor VIII or ELAM-1.

BACKGROUND AND DESIGN: Historically, the Kaposi's sarcoma (KS) tumor cell was generally thought to be derived from endothelial cells. However, by immunohistochemical staining of lesions, many spindle-shaped cells in KS express factor XIIIa, suggesting that they may be related to dermal dendrocytes, a perivascular cell of monocyte/macrophage lineage. To extend our observations on tissue sections of lesions, four different established KS-derived cell lines before and after cytokine exposure, as well as biopsy specimens of KS lesions, were studied for the expression of endothelial cell and monocyte/macrophage antigens. RESULTS: Multipassaged KS tumor cells grew predominantly as spindle-shaped cells. By immunohistochemical staining, all four KS-derived cell cultures were factor XIIIa and intercellular adhesion molecule 1 positive. Three KS-derived cell cultures were CD14 and vascular cell adhesion molecule 1 positive. Expression of factor XIIIa was upregulated by interferon gamma. By polymerase chain reaction (PCR) analysis, expression of CD4 mRNA was also detected in all four KS-derived cell cultures. Expression of these monocyte/macrophage markers (factor XIIIa, CD14, and vascular cell adhesion molecule 1) on KS in vitro was also detected on spindle cells of KS lesions in vivo. Endothelial leukocyte adhesion molecule 1 and factor VIII expression was not detected on KS-derived cells in vitro before or after exposure to interferon gamma or tumor necrosis factor alpha. While cultured KS cells strongly expressed smooth-muscle alpha-actin, no detection was observed on spindle-shaped cells in vivo. Conversely, while cultured KS cells were devoid of CD34, the tissue biopsy specimens contain strongly positive spindle-shaped cells. Our results demonstrate that while KS cell lines express one antigen shared by endothelial cells, ie, vascular cell adhesion molecule 1, they lack several other endothelial cell markers such as factor VIII and endothelial leukocyte adhesion molecule 1, either before or after cytokine activation. CONCLUSIONS: Kaposi's sarcoma cell lines express several antigens linking them to dermal dendrocytes such as factor XIIIa, CD4, and CD14.

Antigens, CD↗

PubMed1993

Factor XIIIA subunit and Crohn's disease.

Factor XIIIA is the active subunit of plasma factor XIII that is responsible for cross linking fibrin into a stable clot. Sixteen patients with Crohn's disease were studied prospectively from relapse (Crohn's disease activity index > 150) into remission. Plasma factor XIIIA concentrations were significantly lower in active disease (median 63 (95% CI 46-72) U/dl) than remission (median 90 (95% CI 60-112) U/dl; p = 0.002). Plasma factor XIIIA concentrations correlated positively with the activity index (p = 0.005) and platelet count (p = 0.003), and negatively with serum albumin (p = 0.006). In five patients with persistent aggressive disease, the factor XIIIA concentration remained below the lower range of normal despite apparent clinical improvement in response to medical treatment. Tissues from three patients who underwent surgical resection during the study were immunostained for factor XIIIA. Gut mucosal and submucosal macrophages stained strongly for factor XIIIA. In one patient, capillary thrombi near superficial mucosal erosions immunostained for factor XIIIA in macroscopically normal mucosa. Similar changes were identified in more severely inflamed sections of intestine from the other two patients. The demonstration of significantly low plasma factor XIIIA concentrations in active Crohn's disease, and the immunostaining of factor XIIIA in capillary thrombi in the bowel wall, suggest that activation of coagulation may be involved in the pathogenesis of Crohn's disease. The plasma factor XIIIA concentration may prove a useful laboratory marker of disease activity.

Adolescent↗

PubMed1999

Site-directed mutagenesis of the calcium-binding site of blood coagulation factor XIIIa.

Blood coagulation factor XIIIa is a calcium-dependent enzyme that covalently ligates fibrin molecules during blood coagulation. X-ray crystallography studies identified a major calcium-binding site involving Asp(438), Ala(457), Glu(485), and Glu(490). We mutated two glutamic acid residues (Glu(485) and Glu(490)) and three aspartic acid residues (Asp(472), Asp(476), and Asp(479)) that are in close proximity. Alanine substitution mutants of these residues were constructed, expressed, and purified from Escherichia coli. The K(act) values for calcium ions increased by 3-, 8-, and 21-fold for E485A, E490A, and E485A,E490A, respectively. In addition, susceptibility to proteolysis was increased by 4-, 9-, and 10-fold for E485A, E490A, and E485A,E490A, respectively. Aspartic acids 472, 476, and 479 are not involved directly in calcium binding since the K(act) values were not changed by mutagenesis. However, Asp(476) and Asp(479) are involved in regulating the conformation for exposure of the secondary thrombin cleavage site. This study provides biochemical evidence that Glu(485) and Glu(490) are Ca(2+)-binding ligands that regulate catalysis. The binding of calcium ion to this site protects the molecule from proteolysis. Furthermore, Asp(476) and Asp(479) play a role in modulating calcium-dependent conformational changes that cause factor XIIIa to switch from a protease-sensitive to a protease-resistant molecule.

Amino Acid Sequence↗