ISOLATION AND CHARACTERIZATION OF THE GENES FOR THE a AND b SUBUNITS OF HUMAN COAGULATION FACTOR XIII

1987 ◽  
Author(s):  
A Ichinose ◽  
R E Bottenus ◽  
K R Loeb ◽  
E W Davie
Keyword(s):  
Amino Acid ◽  
Factor Xiii ◽  
Coagulation Factor ◽  
Amino Acid Sequences ◽  
Binding Region ◽  
Recombinant Phage ◽  
B Subunit ◽  
Isolation And Characterization ◽  
A Subunit ◽  
B Subunits

Factor XIII (plasma transglutaminase, fibrin stabilizing factor) is a plasma protein that plays an important role in the final stages of blood coagulation and fibrinolysis. The molecule occurs in blood as a tetramer (a2b2) consisting of two a. subunits and two b subunits. Recently, we have determined the amino acid sequences for both the a. and b subunits of human factor XIII by a combination of cDNA cloning and amino acid sequence analysis. cDNAs coding for the a (3.8 Kb) and b (2.2 Kb) subunits were used for the screening of human genomic DNA libraries. Among 12 × 106 recombinant phage, ∼30 have been shown to contain the sequences for the a subunit and ∼10 have been shown to contain the gene for the b subunit of factor XIII. The clones coding for the a. subunit span ∼90 Kb and have been characterized by restriction mapping. Southern blotting, and DNA sequencing. Both 5’ and 3’ ends of the genomic clones correspond to the 5’ and 3’portions of the cDNA for the a.subunit of factor XIII. The DNA sequence revealed that the activation peptide released ^thrombin (amino acid residues 137), the first putative Ca2+ binding region (around residue 251), the active Site Cys (amino acid residue 314), and the second putative Ca2+ binding region (around residue 473) are encoded by separate exons. Accordingly, the intervening sequences may separate the a subunit into functional and structural domains. The gene organization for the b subunit will also be presented. (Supported by NIH Grant HL 16919.)

scholarly journals Identification of Potential Novel Interacting Partners for Coagulation Factor XIII B (FXIII-B) Subunit, a Protein Associated with a Rare Bleeding Disorder

2019 ◽  
Vol 20 (11) ◽  
pp. 2682 ◽  
Author(s):  
Sneha Singh ◽  
Mohammad Suhail Akhter ◽  
Johannes Dodt ◽  
Peter Volkers ◽  
Andreas Reuter ◽  
...  
Keyword(s):  
Factor Xiii ◽  
Regulatory Protein ◽  
Coagulation Factor ◽  
Factor H ◽  
Complement C3 ◽  
Complement Factor ◽  
B Subunit ◽  
Coagulation Factor Xiii ◽  
A Subunit ◽  
Complement C1q

Coagulation factor XIII (FXIII) is a plasma-circulating heterotetrameric pro-transglutaminase complex that is composed of two catalytic FXIII-A and two protective/regulatory FXIII-B subunits. FXIII acts by forming covalent cross-links within a preformed fibrin clots to prevent its premature fibrinolysis. The FXIII-A subunit is known to have pleiotropic roles outside coagulation, but the FXIII-B subunit is a relatively unexplored entity, both structurally as well as functionally. Its discovered roles so far are limited to that of the carrier/regulatory protein of its partner FXIII-A subunit. In the present study, we have explored the co-presence of protein excipients in commercial FXIII plasma concentrate FibrogamminP by combination of protein purification and mass spectrometry-based verification. Complement factor H was one of the co-excipients observed in this analysis. This was followed by performing pull down assays from plasma in order to detect the putative novel interacting partners for the FXIII-B subunit. Complement system proteins, like complement C3 and complement C1q, were amongst the proteins that were pulled down. The only protein that was observed in both experimental set ups was alpha-2-macroglobulin, which might therefore be a putative interacting partner of the FXIII/FXIII-B subunit. Future functional investigations will be needed to understand the physiological significance of this association.

scholarly journals The Structure of Blood Coagulation Factor XIII Is Adapted to Oxidation

Biomolecules ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 914
Author(s):  
Alexandra Vasilyeva ◽  
Lyubov Yurina ◽  
Alexander Shchegolikhin ◽  
Maria Indeykina ◽  
Anna Bugrova ◽  
...  
Keyword(s):  
Blood Coagulation ◽  
Factor Xiii ◽  
Critical Role ◽  
Coagulation Factor ◽  
Amino Acid Residues ◽  
B Subunit ◽  
Covalent Crosslinking ◽  
Coagulation Factor Xiii ◽  
A Subunit ◽  
Enzymatic Activation

The blood coagulation factor XIII (FXIII) plays a critical role in supporting coagulation and fibrinolysis due to both the covalent crosslinking of fibrin polymers, rendering them resistant to plasmin lysis, and the crosslinking of fibrin to proteins of the fibrinolytic system. The hypochlorite-mediated oxidation of the blood coagulation factor XIII (FXIII) at the different stages of its enzymatic activation is studied for the first time in this paper. The consolidated results obtained with the aid of MS/MS, electrophoresis, and colorimetry demonstrate that in the process of FXIII’s conversion into FXIIIa, the vulnerability of FXIII to hypochlorite-induced oxidation increased as follows: native FXIII < FXIII + Ca2+ << FXIII + Ca2+/thrombin. The modification sites were detected among all the structural regions of the catalytic FXIII-A subunit, except for the activation peptide, and embraced several sushi domains of the FXIII-B subunit. Oxidized amino acid residues belonging to FXIII-A are surface-exposed residues and can perform an antioxidant role. The regulatory FXIII-B subunits additionally contribute to the antioxidant defense of the catalytic center of the FXIII-A subunits. Taken together, the present data along with the data from previous studies demonstrate that the FXIII proenzyme structure is adapted to oxidation.

Abstract 1653: The Role of the Fibrinogen Alpha Chain in Controlling the Rate of Factor XIII Activation: Implications for Coronary Artery Thrombus Formation

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Kerrie A Smith ◽  
Penelope J Adamson ◽  
Robert A Ariens ◽  
Helen Philippou ◽  
Peter J Grant
Keyword(s):  
Factor Xiii ◽  
Plaque Rupture ◽  
Thrombus Formation ◽  
Coagulation Factor ◽  
Cross Linking ◽  
Active Site Cysteine ◽  
B Subunit ◽  
Thrombin Cleavage ◽  
A Subunit ◽  
Α Chain

Background: The development of an obstructive arterial thrombus is dependent on the generation of a platelet rich fibrin mesh secondary to plaque rupture. Fibrin itself is formed by the cleavage of fibrinogen by thrombin with subsequent fibrin cross-linking by activated Factor XIII. Critical to these processes, fibrinogen α-chain residues 242– 424 have a major regulatory role in the dissociation of coagulation Factor XIII A subunit (FXIIIA) from the Factor XIII B subunit (FXIIIB), however the mechanisms for this enhancing effect have not been determined. Methods: α-Chain residues 242– 424 were expressed as a GST-fusion protein in E .coli, along with six further truncations; αC fragments α242– 402, α242–387, α242–374, α242–341, α242–289 and α242–265. FXIII-A and an inactive double thrombin cleavage mutant (R37A/K513A) were also expressed as GST-fusion proteins in E . coli . Results: Using surface plasmon resonance (SPR) and ELISA we can confirm the presence of at least two binding sites on the αC for activated FXIII-A within residues α388 – 424 and α242–265 with an overall KD of 4.9 ± 2.29 μM. This interaction was specific for activated FXIII-A, as the inactive FXIII-A double thrombin cleavage mutant did not bind to the αC as indicated by SPR and ELISA. We demonstrated that the binding of the αC is dependent on a conformational change which occurs during activation of FXIII-A with calcium and thrombin. FXIII-A activated with thrombin alone did not bind αC as determined by SPR and ELISA. Competition studies showed that the αC competitively inhibits binding with a K I of 1.2 μM confirming the specificity of this interaction. Iodoacetamide blocking of FXIII-A active site cysteine did not prevent binding of the αC suggesting this interaction is independent of FXIII-A cross-linking. Furthermore we have identified a novel interaction between FXIIIB and α-chain by ELISA. Utilising the αC fragments we have localised two binding regions between α290–341 and α242–265 with a K D of 123nM ± 11 and 286nM ± 24.7 respectively. Conclusions: The binding of both FXIII A- and B-subunits to this area of the fibrinogen α-chain suggests a crucial role in controlling the amount of activated FXIII generated for clot stabilisation and presents a potential target for therapeutic intervention.

Coagulation Factor XIII and Cardiovascular Disease in UK Asian Patients Undergoing Coronary Angiography

2001 ◽  
Vol 85 (03) ◽  
pp. 408-411 ◽  
Author(s):  
Michael Mansfield ◽  
Peter Grant ◽  
Darren Warner
Keyword(s):  
Insulin Resistance ◽  
Cardiovascular Disease ◽  
Coronary Angiography ◽  
Factor Xiii ◽  
Coagulation Factor ◽  
Asian Population ◽  
B Subunit ◽  
Asian Patients ◽  
Coagulation Factor Xiii ◽  
The Uk

SummaryPossession of the coagulation factor XIII Val34Leu (FXIIIVal34Leu) polymorphism is associated with protection against myocardial infarction (MI) in Caucasians, in the absence of features of insulin resistance. The role of this polymorphism in the UK Asian population, with its high prevalence of insulin resistance and ischaemic heart disease, is unknown. We investigated the frequency of genotypes at this polymorphism, and measures of circulating FXIII in a group of UK Asians attending for coronary angiography. Genotype at the FXIIIVal34Leu polymorphism was not associated with MI. FXIII B-subunit levels correlated with waist: hip ratio (r = 0.19, p 0.005), HbA1c (r = 0.18, p 0.05), fasting triglycerides (r = 0.21, p 0.005), total cholesterol (r = 0.29, p 0.0005) and PAI-1 antigen (r = 0.24, p 0.005). An association between FXIIIVal34Leu and FXIII cross-linking activity was confirmed in these subjects (one-way ANOVA p 0.0005). This evidence does not support the hypothesis that FXIIIVal34Leu is protective against MI in the UK Asian population. FXIII B-subunit levels are strongly linked to risk factors for cardiovascular disease, suggesting an underlying association with insulin resistance.

Isolation and characterization of some novel genes of the apolipoprotein A-I family in Japanese eel, Anguilla japonica

2011 ◽  
Vol 6 (4) ◽  
pp. 545-557 ◽  
Author(s):  
Malay Choudhury ◽  
Takahiro Oku ◽  
Shoji Yamada ◽  
Masaharu Komatsu ◽  
Keita Kudoh ◽  
...  
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Amino Acid ◽  
Consensus Sequence ◽  
Structural Features ◽  
Lipid Binding ◽  
Japanese Eel ◽  
Amino Acid Sequences ◽  
Novel Genes ◽  
Isolation And Characterization

AbstractApolipoproteins such as apolipoprotein (apo) A-I, apoA-IV, and apoE are lipid binding proteins synthesized mainly in the liver and the intestine and play an important role in the transfer of exogenous or endogenous lipids through the circulatory system. To investigate the mechanism of lipid transport in fish, we have isolated some novel genes of the apoA-I family, apoIA-I (apoA-I isoform) 1–11, from Japanese eel by PCR amplification. Some of the isolated genes of apoIA-I corresponded to 28kDa-1 cDNAs which had already been deposited into the database and encoded an apolipoprotein with molecular weight of 28 kDa in the LDL, whereas others seemed to be novel genes. The structural organization of all apoIA-Is consisted of four exons separated by three introns. ApoIA-I10 had a total length of 3232 bp, whereas other genes except for apoIA-I9 ranged from 1280 to 1441 bp. The sequences of apoIA-Is at the exon-intron junctions were mostly consistent with the consensus sequence (GT/AG) at exon-intron boundaries, whereas the sequences of 3′ splice acceptor in intron 1 of apoIA-I1-7 were (AC) but not (AG). The deduced amino acid sequences of all apoIA-Is contained a putative signal peptide and a propeptide of 17 and 5 amino acid residues, respectively. The mature proteins of apoIA-I1-3, 7, and 8 consisted of 237 amino acids, whereas those of apoIA-I4-6 consisted of 239 amino acids. The mature apoIA-I10 sequence showed 65% identity to amino acid sequence of apoIA-I11 which was associated with an apolipoprotein with molecular weight of 23 kDa in the VLDL. All these mature apoIA-I sequences satisfied the common structural features depicted for the exchangeable apolipoproteins such as apoA-I, apoA-IV, and apoE but apoIA-I11 lacked internal repeats 7, 8, and 9 when compared with other members of apoA-I family. Phylogenetic analysis showed that these novel apoIA-Is isolated from Japanese eel were much closer to apoA-I than apoA-IV and apoE, suggesting new members of the apoA-I family.

The gene for coagulation factor XIII a subunit (F13A) is distal to HLA on chromosome 6

1984 ◽  
Vol 67 (4) ◽  
pp. 406-408 ◽  
Author(s):  
P. G. Board ◽  
M. Reid ◽  
S. Serjeantson
Keyword(s):  
Factor Xiii ◽  
Coagulation Factor ◽  
Chromosome 6 ◽  
Coagulation Factor Xiii ◽  
A Subunit

scholarly journals GyrB mutations in Staphylococcus aureus strains resistant to cyclothialidine, coumermycin, and novobiocin.

1996 ◽  
Vol 40 (4) ◽  
pp. 1060-1062 ◽  
Author(s):  
M Stieger ◽  
P Angehrn ◽  
B Wohlgensinger ◽  
H Gmünder
Keyword(s):  
Staphylococcus Aureus ◽  
Binding Mode ◽  
Atp Binding ◽  
Subunit Gene ◽  
Binding Region ◽  
B Subunit ◽  
B Subunits ◽  
Atp Binding And Hydrolysis

The sequence of the gyrase B subunit gene from Staphylococcus aureus strains resistant to the gyrase B subunit inhibitors cyclothialidine, coumermycin, and novobiocin has been determined. The residues altered in the resistant gyrase B subunits map to the ATP-binding region, suggesting that the drugs inhibit ATP binding and hydrolysis. The pattern of cross-resistances indicates that the detailed binding mode of the compounds differs.

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.

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