Amino acid sequence of the peptide released from bovine factor XIII following activation by thrombin

Nakamura, Shin; Iwanaga, Sadaaki; Suzuki, Tomohiko; Mikuni, Yuko; Konishi, Kazuhiko

doi:10.1016/0006-291x(74)90919-x

Cited by 28 publications

(12 citation statements)

References 9 publications

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“…1). The amino end group of placental FXIIIa is blocked; presumably the amino-terminal serine is N-acetylated, as it is in human plasma and platelet FXIIIa (6) and in bovine plasma FXIIIa (7). The amino acid composition deduced from the protein sequence (Table 1) agrees well with that of Bohn and Schwick (8).…”

Section: Methodssupporting

confidence: 78%

“…The b chain has no role for FXIIIa activity, but it is thought to have a protective function for circulating plasma FXIII. Although composed only of a chains, the platelet and placental enzymes (FXIIIa) are activated by thrombin and Ca2+ in a manner similar to that for the plasma FXIII zymogen (1)(2)(3)(4)(5)(6)(7)(8)(9)(10). Results described here indicate that human FXIIIa molecules derived from plasma, platelets, and placenta are apparently identical in primary structure.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Primary structure of blood coagulation factor XIIIa (fibrinoligase, transglutaminase) from human placenta.

Takahashi¹,

Takahashi²,

Putnam³

1986

Proc. Natl. Acad. Sci. U.S.A.

148

114

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We have determined the primary structure of human placental factor XIJ1a, an enzyme [fibrinoligase, transglutaminase, fibrin-stabilizing factor, EC 2.3.2.13 (protein-glutamine:amine r-glutamyltransferase)] that forms intermolecular isopeptide bonds between fibrin molecules as the last step in blood coagulation. Placental factor XIa is an unglycosylated polypeptide chain of 730 amino acid residues (Mr = 83,005) that appears to be identical to the a subunit of the plasma zymogen factor XI. Ca2+-dependent activation of factor XHIa by thrombin removes a blocked amino-terminal peptide and unmasks a reactive thiol group at Cys-314. A second specific cleavage after Lys-513 by thrombin inactivates factor XIIa and produces an amino-terminal 56-kDa fragment and a 24-kDa fragment. The amino acid sequence of factor XIIa is unique and does not exhibit internal homology, but its active center is similar to that of the thiol proteases. The probable Ca2+-binding site of factor XIa has been identified by homology to the high-affmity sites in calmodulins. Knowledge of the primary structure of factor XMa will aid elucidation of the mechanism of its enzymatic action and that of the many tissue transglutaminases ofwhich it is the prototype. This will also facilitate production of factor XIa by recombinant DNA technology for use in treatment of congenital factor XIII deficiencies and in the postoperative healing of wounds.Factor XIIIa [FXIIIa; fibrinoligase, fibrin-stabilizing factor, EC 2.3.2.13 (protein-glutamine:amine y-glutamyltransferase)] is the last enzyme generated in the blood coagulation cascade (1, 2). It is a transglutaminase or transamidating enzyme that forms intermolecular y-glutamyl-e-lysine crosslinks between fibrin molecules, thereby stabilizing the fibrin clot mechanically and also conferring resistance to proteolysis. FXIIIa has a cysteine thiol active site; however, unlike thiol proteases such as cathepsins B and H that cleave peptide bonds, FXIIIa forms intermolecular isopeptide bridges in fibrin. Other substrates for FXIIIa include coagulation factor V, a2-macroglobulin, platelet myosin, actin, and fibronectin. Thus, in addition to its essential role in blood clotting, FXIIIa may function to stabilize cell surface molecules and membranes. FXIIIa is the prototype of a class of Ca2+-dependent transglutaminases with thiol active centers that are widespread in animal tissues and have been associated with cell proliferation, embryonic development, and growth (3-5). However, none except FXIIIa has been puri-, fled and characterized, and until now little has been published about the structure of FXIIIa except the sequence of the amino-terminal activation peptide of the human (6) and bovine (7) plasma FXIIIa. We report here the primary structure of human placental FXIIIa, including the sequence at the active site, and the sites of activation and inactivation by thrombin and the probable Ca2`-binding site.

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Section: Methodssupporting

confidence: 78%

mentioning

confidence: 99%

Primary structure of blood coagulation factor XIIIa (fibrinoligase, transglutaminase) from human placenta.

Takahashi¹,

Takahashi²,

Putnam³

1986

Proc. Natl. Acad. Sci. U.S.A.

148

114

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“…From the deduced amino acid sequence, the mature (16,29,31). This is in agreement with an analysis of 1764 eukaryotic protein sequences that showed that serine, alanine, or glycine in the second position favor removal ofthe initiator methionine and N-acetylation of the NH2-terminal amino acid (32).…”

Section: Resultssupporting

confidence: 65%

Characterization of cDNA coding for human factor XIIIa.

Grundmann¹,

Amann²,

Zettlmeissl³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

119

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A cDNA library prepared from human placenta has been screened for sequences coding for factor XIIIa, the enzymatically active subunit of the factor Xm complex that stabilizes blood clots through crosslinking of fibrin molecules. Two oligonucleotides, based on the amino acid sequences of tryptic peptides of factor XMa, were used as hybridization probes. Of 0.36 x 106 independent recombinants, 1 clone was identified that hybridized to both probes. The insert of 1704 base pairs coded for the amino-terminal 541 amino acid residues of the mature factor XIIIa molecule. Blot-hybridization analysis using this cDNA as a probe showed that the factor XIIIa mRNA from placenta has a size of approximately 4000 bases. The insert was used to rescreen cDNA libraries and to identify further factor XMa-specific. sequences. The total length of the isolated factor XILIa cDNA is 3905 bases, and it codes for a protein of 732 amino acids. In spite of the presence of factor XIII in blood plasma, we could not identify a leader sequence typical for secreted proteins.

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“…This is in keeping with the fact that the physiological substrates of thrombin, i.e. prothrombin and factor XIII, also contain Pro at Pz [7,8], and studies with sequence-analogous inhibitors also support this view [16,26,29]. This is reflected in the respective contributions in both substrate series.…”

Section: Regression Analysis Of Kinetic Datasupporting

confidence: 59%

Study of the Specificity of Thrombin with Tripeptidyl-p-nitroanilide Substrates

Pozsgay

Szabó

Elödi

et al. 2005

European Journal of Biochemistry

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The kinetic behaviour of human thrombin has been studied with 26 tripeptidyl-p-nitroanilide substrates protected at the N terminus and with 9 unprotected ones. By the regression analysis of experimentally determined I/&,, k,,, and /cCat/Km values the individual contribution of each side chain of the various substrates to the kinetic parameters was calculated.The contributions to the kinetic parameters of the best substrates provide information about the structure of the binding site. The interaction of subsites S1 and PI, which determines primary specificity, proved to be marginal on the basis of contribution values, though it depends upon this contact whether the substrate is hydrolyzed at all. At subsite Sz proline appeared to be favourable. Subsite S3 plays an important role in efficiency. The best parameters were obtained here with the D configurations of bulky amino acid residues. The aromatic protecting groups applied did not improve the properties of substrates. BzDPhe-Pro-Arg-Nan was predicted by calculation to be better than the protected substrates assayed. The compound was synthesized and tested. Its experimentally determined l/K,,,, 55.1 mM-', was in good agreement with 50.9 mM-' found by calculation.Several serine proteases, e.g. thrombin, trypsin, plasmin and factor Xa, hydrolyze peptide bonds at the carboxylic group of basic amino acids. Their sensitivities to substrate specificity are, however, different. Among them thrombin is the most specific which, apart from a few exceptions, splits next to arginine [1,2] [9] and growth hormones [lo]. According to Muszbek [ I l l in actin it also hydrolyzes next to lysine, whereas hydrolysis does not occur next to ornithine [12].The detailed structure of the substrate-binding sites of thrombin is not yet known. The investigations on the specificity and substrate-binding sites have so far been carried out, in addition to fibrinogen, with peptide derivatives analogous to fibrinogen sequences [13,14] [2,16] and similar hydrophobic side chains, whereas results concerning the S3 substrate-binding subsite are contradictory. Scheraga holds that S3 is rather narrow; it is conceivable, though, that more than one binding domain is operative against long peptides [12].In an attempt to study the substrate-binding sites of thrombin we applied a mathematical method [19], by the aid of which, through the regression analysis of the kinetic constants measured for various tripeptidyl-p-nitroanilide substrates, the interaction between substrate side chains and the

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Amino acid sequence of the peptide released from bovine factor XIII following activation by thrombin

Cited by 28 publications

References 9 publications

Primary structure of blood coagulation factor XIIIa (fibrinoligase, transglutaminase) from human placenta.

Primary structure of blood coagulation factor XIIIa (fibrinoligase, transglutaminase) from human placenta.

Characterization of cDNA coding for human factor XIIIa.

Study of the Specificity of Thrombin with Tripeptidyl-p-nitroanilide Substrates

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