The binding of divalent metal ions to platelet factor XIII modulates its proteolysis by trypsin and thrombin

Mary, Ann; Achyuthan, Komandoor E.; Greenberg, Charles S.

doi:10.1016/0003-9861(88)90110-5

Cited by 27 publications

(48 citation statements)

References 27 publications

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“…When transglutaminase activity was induced by I M-NaCl a saturating concentration for CaCI2 was reached at around 20 mm, and even at 1 mm 600% of the maximal activity could be obtained. The observed dependence of I M-NaCl-induced activation of placenta FXIII on Ca2+ concentration is quantitatively well comparable with that of thrombin activation of platelet FXIII [23]. Un a low but reproducible transglutaminase activity.…”

Section: Resultssupporting

confidence: 71%

Non-proteolytic activation of cellular protransglutaminase (placenta macrophage factor XIII)

Polgár

Hidasi

Muszbek

1990

Biochemical Journal

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Plasma Factor XIII is a zymogen (plasma protransglutaminase) with the tetrametric structure A2B2, whereas the cellular protransglutaminase, i.e. Factor XIII in the platelet and monocyte/macrophage, consists exclusively of A subunits (A2). It is generally accepted that at Ca2+ concentrations comparable with that in plasma the proteolytic removal of an Nterminal activation peptide is the prerequisite for the Ca2+-induced formation of a catalytically active configuration of subunit A. In this study it was demonstrated that at high concentrations NaCl or KCI induced a non-proteolytic activation of cellular (placental macrophage) but not plasma protransglutaminase. The activation depended on time and salt concentration, and Ca2 , in the range 0-20 mm, greatly enhanced the activation process. At 1.25 M-NaCl maximal activation occurred within 60 min in the presence of 2 mM-CaC12, and even at physiological NaCl concentration a slow progressive activation could be observed in the presence of Ca2+. The specific activity of salt-activated Factor XIII was 1.5-2.0-fold higher than that obtained after thrombin activation. The non-proteolytic activation of cellular protransglutaminase was abolished by the addition of subunit B of plasma Factor XIII in stoichiometric amount, which suggests that (one of) the physiological function(s) of the B subunit in plasma Factor XIII is to prevent the slow spontaneous activation of A subunit that would occur in a plasmatic environment. INTRODUCTIONFactor XIII (FXIII) of blood coagulation present in the plasma is a zymogen (plasma protransglutaminase) of tetrameric structure (A2B2). The active site formed in the course of an activation process is located on the A subunit while subunit B remains enzymically inactive. The presence of FXIII has also been verified in platelets [1], monocytes and monocyte-derived macrophages [2][3][4][5][6][7]. In contrast with the plasma FXIII, however, this cellular protransglutaminase consists exclusively of A subunits (A2). The active transglutaminase (FXIIIa) formed from FXIII catalyses an acyl-transfer reaction in which the carboxamide group of a peptide-bound glutamine residue is the acyl

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Section: Resultssupporting

confidence: 71%

Non-proteolytic activation of cellular protransglutaminase (placenta macrophage factor XIII)

Polgár

Hidasi

Muszbek

1990

Biochemical Journal

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“…There is further in vitro evidence that calcium and other divalent cations have effects on proteolytic susceptibility (22) and heat stability (23). Several lanthanide ions have been shown to replace calcium ions during fXIII activation (10 -40 M).…”

mentioning

confidence: 99%

Identification of the Calcium Binding Site and a Novel Ytterbium Site in Blood Coagulation Factor XIII by X-ray Crystallography

Fox

Yee

Pedersen

et al. 1999

Journal of Biological Chemistry

110

124

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The presence or absence of calcium determines the activation, activity, oligomerization, and stability of blood coagulation factor XIII. To explore these observed effects, we have determined the x-ray crystal structure of recombinant factor XIII A 2 in the presence of calcium, strontium, and ytterbium. The main calcium binding site within each monomer involves the main chain oxygen atom of Ala-457, and also the side chains from residues Asn-436, Asp-438, Glu-485, and Glu-490. Calcium and strontium bind in the same location, while ytterbium binds several angstroms removed. A novel ytterbium binding site is also found at the dimer two-fold axis, near residues Asp-270 and Glu-272, and this site may be related to the reported inhibition by lanthanide metals (Achyuthan, K. E., Mary, A., and Greenberg, C. S. (1989) Biochem. J. 257, 331-338). The overall structure of ion-bound factor XIII is very similar to the previously determined crystal structures of factor XIII zymogen, likely due to the constraints of this monoclinic crystal form. We have merged the three independent sets of water molecules in the structures to determine which water molecules are conserved and possibly structurally significant.The biological importance of factor XIII (fXIII) 1 (EC 2.3.2.13) lies in its ability to form new covalent bonds between protein chains. This activity was first recognized while studying blood coagulation; fXIII was required to form an insoluble clot (1). The activated form of fXIII covalently cross-links two fibrin molecules via an isopeptide bond between the side chains of a glutamine and a lysine located in the C-terminal region of the ␥-chain. Over a longer time period in coagulation, it also forms cross-links between the ␣-and ␥-chains of fibrin (2), and between ␣ 2 -anti-plasmin and fibrin (3). fXIII has been shown to react with more than fibrin (4), and it has recently been found in brain tumors (5) and arthritic joints (6), and there are cases of fXIII deficiencies (7,8). Factor XIII is a member in the family of transglutaminases (TGases), which have a wide range of biological functions (9).Like most coagulation factors, fXIII is synthesized as a zymogen and then cleaved by a protease to become an active enzyme. The structure of fXIII zymogen was determined several years ago (10, 11). In this crystal form, the active site cysteine, Cys-314, is inaccessible to solvent and is not available for catalysis.Physiologically, calcium ions are required for fXIII activation and for TGase activity. In the blood, activation of circulating fXIII requires thrombin cleavage, calcium ions (1.5 mM) (12-14), and fibrin(ogen) (15). High levels of calcium (Ͼ50 mM) can activate fXIII without the use of thrombin (15), and it has recently been shown that platelet fXIII can be activated nonproteolytically in vivo (16).Based on the amino acid sequence, the calcium binding site was predicted to be in a region (residues 468 -479) with high similarity to the EF-hand motif (17, 18). The main calcium binding site, as seen in the preliminary cryst...

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“…The activation of platelet FXIII occurs through a thrombin cleavage of the Arg37-Gly38 peptide bond near the N-terminus of the a-chains [2]. Thrombin and trypsin have been reported both to activate and to inactivate FXIII [3][4][5][6][7]. It was reported that Ca(II) and several other bivalent metal ions played a crucial role during the proteolytic activation of FXIII and in preservation of the transglutaminase activity of FXIIIa [7,8].…”

Section: Introductionmentioning

confidence: 99%

“…Factor XIIIa formed by thrombin or trypsin in the presence of 40 ,uM-Tb(III) ions, however, was indistinguishable from Factor XIIIa formed in the presence of 2-5 mMCa(II) ions with respect to molecular mass and transglutaminase activity. [7,8]. In the absence of metal ions (5 mM-EDTA), trypsin and thrombin treatment resulted in rapid inactivation of FXIII [6,7].…”

mentioning

confidence: 99%

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Tb(III)-ion-binding-induced conformational changes in platelet factor XIII

Achyuthan

Mary

Greenberg

1989

Biochemical Journal

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Ca(II) ions are crucial during proteolytic conversion of Factor XIII zymogen into the active enzyme Factor XIIIa. Factor XIII proteolysed by thrombin or trypsin in the presence of 5 mM-EDTA resulted in rapid inactivation of transglutaminase activity. Factor XIIIa formed by thrombin or trypsin in the presence of 40 ,uM-Tb(III) ions, however, was indistinguishable from Factor XIIIa formed in the presence of 2-5 mMCa(II) ions with respect to molecular mass and transglutaminase activity. [7,8]. In the absence of metal ions (5 mM-EDTA), trypsin and thrombin treatment resulted in rapid inactivation of FXIII [6,7].

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The binding of divalent metal ions to platelet factor XIII modulates its proteolysis by trypsin and thrombin

Cited by 27 publications

References 27 publications

Non-proteolytic activation of cellular protransglutaminase (placenta macrophage factor XIII)

Non-proteolytic activation of cellular protransglutaminase (placenta macrophage factor XIII)

Identification of the Calcium Binding Site and a Novel Ytterbium Site in Blood Coagulation Factor XIII by X-ray Crystallography

Tb(III)-ion-binding-induced conformational changes in platelet factor XIII

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