Calcium and thiol reactivity of human plasma clotting factor XIII

Cooke, R. D.; Pestell, Timothy C.; Holbrook, J. John

doi:10.1042/bj1410675

Cited by 16 publications

(9 citation statements)

References 32 publications

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“…Calcium appears to play an intriguing regulatory role in FXIII [19–20, 50–51]. Low mM levels already begin to initiate important conformational changes needed to expose the active site and make the enzyme ready to accommodate incoming substrates.…”

Section: Discussionmentioning

confidence: 99%

Role of calcium in the conformational dynamics of factor XIII activation examined by hydrogen–deuterium exchange coupled with MALDI-TOF MS

Woofter¹,

Maurer²

2011

Archives of Biochemistry and Biophysics

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Factor XIII catalyzes formation of γ-glutamyl-ε-lysyl crosslinks within fibrin clots. FXIII A2 can be activated proteolytically with thrombin and low mM Ca2+ or nonproteolytically with high monovalent/divalent cations along with low mM Ca2+. Physiologically, FXIII A2 is poised to respond to transient influxes of Ca2+ in a Na+ containing environment. A successful strategy to monitor FXIII conformational events is hydrogen-deuterium exchange (HDX) coupled with mass spectrometry. FXIII A2 was examined in the presence of different cations (Ca2+, Mg2+, Ba2+, Cu2+, Na+, TMAC+, and EDA2+) ranging from 1–2 mM, physiological Ca2+ concentration, to 50–500mM for nonproteolytic activation. Increases in FXIII solvent exposure could already be observed at 1 mM Ca2+ for the dimer interface, the catalytic site, and glutamine substrate regions. By contrast, solvent protection was observed at the secondary cleavage site. These events occurred even though 1mM Ca2+ is insufficient for FXIII activation. The metals 1mM Mg2+, 1mM Ba2+, and 1mM Cu2+ each led to conformational changes, many in the same FXIII regions as Ca2+. FXIII could also be activated nonproteolytically with 500mM tetramethylammonium chloride (TMAC+) and 500mM ethylenediamine (EDA2+), both with 2mM Ca2+. These different HDX studies help reveal the first FXIII segments that respond to physiological Ca2+ levels.

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Section: Discussionmentioning

confidence: 99%

Role of calcium in the conformational dynamics of factor XIII activation examined by hydrogen–deuterium exchange coupled with MALDI-TOF MS

Woofter¹,

Maurer²

2011

Archives of Biochemistry and Biophysics

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“…In addition to chelating CaZ+ with ethelenediaminetetraacetic acid disodium salt (EDTA) or ATP, or inactivating the enzyme by Zn2+ [ 161 , mercurials, or disulfides [17] , the above type of formulation of the pathway suggests that primary aminesif they satisfy the specificity requirements of the enzymemight interfere with the formation of the P-CONH-P' product. Indeed, we found that such synthetic compounds could inhibit cross-linking rather effectively even in the complex biological system of clotting in whole blood [13, 181.…”

Section: Transamidase-catalyzed Cross-linking Of Proteinsmentioning

confidence: 99%

Formation of γ‐glutamyl‐ϵ‐lysine bridges between membrane proteins by a Ca²⁺‐regulated enzyme in intact erythrocytes

1978

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A rise in the intracellular concentration of Ca2+-ions in human erythrocytes causes the formation of high-molecular-weight membrane protein polymers, cross-linked by gamma-glutamyl-epsilon-lysine side chain bridges. Cross-linking involves proteins at the cytoplasmic side of the membrane (band 4.1, spectrin, and band 3 materials) and the reaction is catalyzed by the intrinsic transglutaminase. This enzyme is regulated by Ca2+-ions and it exits in a latent form in normal cells. The protein polymer, isolated from the membranes of Ca2+-loaded intact human red cells, is heterogeneous in size and may contain as many as 6 moles of gamma-glutamyl-epsilon-lysine cross-links per 100,000 gm of protein. Synthetic compounds, which either compete against the epsilon-lysine cross-linking functionalities of the protein substrates (eg, histamine, aminoacetonitrile, cystamine) or directly inactivate the transamidase (eg, cystamine), inhibit the membrane polymerization reaction in intact human erythrocytes. They also interfere with the Ca2+-induced irreversible shape change from discocyte to echinocyte and inhibit the irreversible loss of membrane deformability. Thus, the transamidase-catalyzed production of gamma-glutamyl-epsilon-lysine cross-links in the membrane may be a common denominator in these cellular manifestations.

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“…Ca2+ causes Factor XIIIa to dissociate into sub-units, the a' subunits subsequently aggregating into a misty precipitate (Cooke & Holbrook, 1974b), and is also required for inhibition of the essential thiol of plasma Factor XIII (Cooke et al, 1974). The Ca2+ concentrations required for both these processes are very much higher than that required to observe maximum assay rates, indicating that they may be non-physiological reactions.…”

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confidence: 99%

Calcium-induced dissociation of human plasma factor XIII and the appearance of catalytic activity

Cooke

1974

Biochemical Journal

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1. The Ca(2+) dependence of the activity of plasma Factor XIII(a) was studied by using the continuous assay based on the incorporation of dansylcadaverine into dephosphorylated acetylated beta-casein (beta-substrate). The K(m) for Ca(2+) is about 0.170mm. 2. At low concentrations of Ca(2+) there was a lag in attaining the steady-state rate. The size of the lag was decreased and eventually abolished if the enzyme was preincubated with a high concentration of Ca(2+) before assay. The concentration of Ca(2+) required to decrease the lag phase by 50% in 10min depended on the protein concentration: at 0.87mg of protein/ml it required 17mm-Ca(2+) and at 0.44mg/ml it needed 10mm-Ca(2+). 3. The concentrations of Ca(2+) required either to abolish the lag phase in the appearance of enzyme activity or to activate the essential thiol for reaction with 5,5'-dithiobis-(2-nitrobenzoate) in 10min incubation were similar at the same protein concentration. This indicated that Ca(2+) induces a conformation change that is responsible for both phenomena. A model is proposed that links this conformation change to the dissociation of the tetrameric enzyme. 4. This was supported by the observation that the addition of excess of b chains to the Factor XIII(a) (a'(2)b(2)) increased the concentration of Ca(2+) required to expose the reactive thiol, and inhibited the Ca(2+)-dependent aggregation of a' chains. 5. Platelet Factor XIII(a) (a'(2)) was inhibited by 5,5'-dithiobis-(2-nitrobenzoate) in the absence of Ca(2+), and no lag phases were observed in attaining the steady-state rate at low Ca(2+) concentrations, thus confirming the model for the activation of the plasma enzyme. 6. The Ca(2+) dependence of platelet Factor XIII(a) indicated that Ca(2+) has an additional role in the enzyme mechanism of the plasma enzyme, perhaps being involved in substrate binding. 7. The dependence of the stability of plasma Factor XIII(a) on Ca(2+) and protein concentration indicates that the decay in activity is related to the tetramer dissociation. 8. beta-Substrate decreased the Ca(2+) concentration required for (1) abolition of the lag phase and (2) enzyme inhibition by thiol reagents. The effect on the former is greater than on the latter. 9. The role of the b chains of the plasma Factor and the evolutionary significance of the plasma and platelet Factors are considered.

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Calcium and thiol reactivity of human plasma clotting factor XIII

Cited by 16 publications

References 32 publications

Role of calcium in the conformational dynamics of factor XIII activation examined by hydrogen–deuterium exchange coupled with MALDI-TOF MS

Role of calcium in the conformational dynamics of factor XIII activation examined by hydrogen–deuterium exchange coupled with MALDI-TOF MS

Formation of γ‐glutamyl‐ϵ‐lysine bridges between membrane proteins by a Ca²⁺‐regulated enzyme in intact erythrocytes

Calcium-induced dissociation of human plasma factor XIII and the appearance of catalytic activity

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Calcium and thiol reactivity of human plasma clotting factor XIII

Cited by 16 publications

References 32 publications

Role of calcium in the conformational dynamics of factor XIII activation examined by hydrogen–deuterium exchange coupled with MALDI-TOF MS

Role of calcium in the conformational dynamics of factor XIII activation examined by hydrogen–deuterium exchange coupled with MALDI-TOF MS

Formation of γ‐glutamyl‐ϵ‐lysine bridges between membrane proteins by a Ca2+‐regulated enzyme in intact erythrocytes

Calcium-induced dissociation of human plasma factor XIII and the appearance of catalytic activity

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Formation of γ‐glutamyl‐ϵ‐lysine bridges between membrane proteins by a Ca²⁺‐regulated enzyme in intact erythrocytes