Thrombin inhibits the anti-myeloperoxidase and ferroxidase functions of ceruloplasmin: relevance in rheumatoid arthritis

Sokolov, A. V.; Acquasaliente, Laura; Kostevich, Valeria A.; Frasson, Roberta; Захарова, Е. М.; Pontarollo, Giulia; Vasilyev, V. B.; Filippis, Vincenzo De

doi:10.1016/j.freeradbiomed.2015.05.016

Cited by 41 publications

(51 citation statements)

References 66 publications

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“…thrombin, serprocidins, and matrix metalloproteinases 2 and 12 (Sokolov et al 2007(Sokolov et al , 2009b(Sokolov et al , 2010a(Sokolov et al , 2015a. Plasma concentration of the socalled ''free copper'' grows in Alzheimer's disease concommittantly with the proteolytic degradation of CP (Squitti et al 2008).…”

Section: Discussionmentioning

confidence: 99%

Interaction of macrophage migration inhibitory factor with ceruloplasmin: role of labile copper ions

et al. 2015

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Macrophage migration inhibitory factor (MIF), a pro-inflammatory cytokine, is a target for pharmacological treatment of sepsis and malignant tumors. Inhibition of tautomerase activity of MIF in reaction with p-hydroxyphenylpyruvate (HPP) was observed in the presence of ceruloplasmin (CP), a copper-containing plasma protein. Binding labile copper ions to CP (CP+Cu(II)) is a prerequisite for MIF inhibiting. CP+Cu(II) is shown to be an uncompetitive inhibitor of MIF (Ki ~ 37 nM), which suggests formation of a complex 'MIF-HPP-CP-Cu(II)'. Filtration of CP+Cu(II) on a column with Chelex-100, otherwise the presence of high concentrations of histidine, cysteine or methionine abrogated the inhibitory effect of CP. Adding salts of Co(II) and Ni(II) that replace copper ions in the labile sites prevented the inhibitory effect of CP+Cu(II). Limited proteolysis of CP by thrombin diminished its oxidase activity in reaction with p-phenylenediamine, but endowed it with the capacity of inhibiting MIF. Covalent modification of MIF by phenylmethylsulfonyl fluoride (PMSF) resulted in binding of MIF-PMSF to CP immobilized on CM5 chip, the dissociation constant being 4.2 μM. In D-galactosamine-sensitized mice CP+Cu(II) increased the LPS-induced lethality from 54 to 100%, while administration of antibodies against MIF prevented the lethal effect. The enhancement by CP+Cu(II) of the pro-inflammatory signal of MIF is discussed.

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Section: Discussionmentioning

confidence: 99%

Interaction of macrophage migration inhibitory factor with ceruloplasmin: role of labile copper ions

et al. 2015

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“…Blotting was performed at 4°C in 10 mM CAPS, pH 10.5, at 50 V for 3 h. Alternatively, the gel bands were excised and subjected to in situ peptide mass fingerprint with trypsin as described (72). After extraction of the tryptic peptides from the gel, the proteolytic mixture was loaded onto a Grace-Vydac (Hesperia, CA) C4 (1 ϫ 10 mm) microbore column connected to an Agilent (Santa Clara, CA) 1290 UHPLC system.…”

Section: Identification Of the Subtilisin Cleavage Sites On Prot And mentioning

confidence: 99%

“…The NeutrAvidin-coated chip was then loaded for 700 s at a flow rate of 10 l/min with a solution (100 l, 0.4 M) of the synthetic peptide Biotinyl-PEG-GpIb␣(268 -282). Notably, NeutrAvidin (pI ϭ 6.3) was used as an excellent alternative to avidin (pI ϭ 10) for minimizing non-specific binding (53,72) Finally, aliquots (100 l) of ␣T and Pre2 solutions at increasing concentrations were injected over the NeutrAvidin sensor chip saturated with biotinyl-PEG-GpIb␣(268 -282) (supplemental Fig. S1) at a flow rate of 10 l/min, with a contact time of 350 s, in 10 mM HEPES, pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.05% polyoxyethylene sorbitan (HBS-EP ϩ ).…”

Section: Sprmentioning

confidence: 99%

Non-canonical proteolytic activation of human prothrombin by subtilisin from Bacillus subtilis may shift the procoagulant–anticoagulant equilibrium toward thrombosis

Pontarollo

Acquasaliente

Peterle

et al. 2017

Journal of Biological Chemistry

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Blood coagulation is a finely regulated physiological process culminating with the factor Xa (FXa)-mediated conversion of the prothrombin (ProT) zymogen to active ␣-thrombin (␣T). In the prothrombinase complex on the platelet surface, FXa cleaves ProT at Arg-271, generating the inactive precursor prethrombin-2 (Pre2), which is further attacked at Arg-320 -Ile-321 to yield mature ␣T. Whereas the mechanism of physiological ProT activation has been elucidated in great detail, little is known about the role of bacterial proteases, possibly released in the bloodstream during infection, in inducing blood coagulation by direct proteolytic ProT activation. This knowledge gap is particularly concerning, as bacterial infections are frequently complicated by severe coagulopathies. Here, we show that addition of subtilisin (50 nM to 2 M), a serine protease secreted by the non-pathogenic bacterium Bacillus subtilis, induces plasma clotting by proteolytically converting ProT into active Pre2, a nicked Pre2 derivative with a single cleaved Ala-470 -Asn-471 bond. Notably, we found that this non-canonical cleavage at Ala-470 -Asn-471 is instrumental for the onset of catalysis in Pre2, which was, however, reduced about 100 -200-fold compared with ␣T. Of note, Pre2 could generate fibrin clots from fibrinogen, either in solution or in blood plasma, and could aggregate human platelets, either isolated or in whole blood. Our findings demonstrate that alternative cleavage of ProT by proteases, even by those secreted by non-virulent bacteria such as B. subtilis, can shift the delicate procoagulant-anticoagulant equilibrium toward thrombosis.Blood coagulation is a finely regulated physiological process that culminates with the factor Xa-mediated conversion of the prothrombin (ProT) 3 zymogen to the active ␣-thrombin (␣T) enzyme, which in turn is responsible for the generation of insoluble fibrin and activation of platelets via the GpIb␣-PAR1 pathway (1, 2). ProT (ϳ72 kDa) is a vitamin K-dependent glycoprotein produced in the liver and circulating at a relatively high plasma concentration (0.1 mg/ml) (3). The domain architecture of ProT includes a Gla domain (residues 1-46), a kringle-1 (residues 65-143) and a kringle-2 (residues 170 -248) domain, and a chymotrypsin-like protease domain (residues 285-579) connected by three intervening linker regions (Lnk-1, -2, and -3) (4). Isolated factor Xa (FXa) has low intrinsic ProTconverting activity, but when it is assembled in the presence of Ca 2ϩ with cofactor Va in the prothrombinase complex on the platelet surface, its ability to activate ProT is increased by about 5 orders of magnitude (5). FXa cleaves ProT in a concerted manner at two sites, i.e. Arg-271 and Arg-320, but the order of peptide bond cleavage is highly context-dependent. On the platelet surface, FXa first cleaves ProT at Arg-271 generating the inactive precursor prethrombin-2 (Pre2), which is attacked by FXa at Arg-320 to generate the active ␣T species, formed by the polypeptide chains Thr-272-Arg-320 and Ile-321-Glu-579 (6). ...

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“…Препарат церулоплазмина (CP) выделяли из цитратной плазмы при помощи ионообменной хроматографии (UNOsphere TM Q, Bio-Rad, США) и аффинной хроматографии (неомицин-агароза). Следы протромбина и тромбина удаляли путем фильтрации препарата через колонки с гепарин-и бензамидин-агарозой [9].…”

Section: материалы и методыunclassified

Celestine blue B as a sensor for hypochlorous acid and HOCL-modified proteins registration

Луценко

Евгеньевна²,

Grigorieva

et al. 2019

Medical academic journal

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Objective — the study of hypochlorous acid (HOCl) and its derivatives production, which catalyzed by human neutrophil myeloperoxidase, using “turn-on” fluorescent sensor — celestine blue B. Materials and methods. Neutrophils were isolated from the venous blood of healthy donors. Phorbol 12-myristate 13-acetate, N-formyl-methionyl-leucyl-phenylalanine, plant lectins, HOCl-modified proteins were used as agonists. N-acetylcysteine, 4-aminobenzoic acid hydrazide, isoniazid and ceruloplasmin were used as regulators of neutrophil myeloperoxidase activity and/or HOCl scavengers. Results. Using a wide range of agonists and inhibitors, it has been shown that celestine blue B is oxidized in vitro by HOCl and its derivatives as a result of neutrophil myeloperoxidase activity. The oxidation of celestine blue B by HOCl-modified human serum albumin (HSA-Cl) and inhibition of this process by monoclonal antibody against HSA-Cl (IgM class) was also found. Conclusion. Based on the developed method using celestine blue B, it is possible to conduct a sensitive analysis for the presence of HOCl-modified proteins (chloramines, etc.), to investigate the effect of various agonists and drugs on myeloperoxidase activity and exocytosis from the neutrophil granules.

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Thrombin inhibits the anti-myeloperoxidase and ferroxidase functions of ceruloplasmin: relevance in rheumatoid arthritis

Cited by 41 publications

References 66 publications

Interaction of macrophage migration inhibitory factor with ceruloplasmin: role of labile copper ions

Interaction of macrophage migration inhibitory factor with ceruloplasmin: role of labile copper ions

Non-canonical proteolytic activation of human prothrombin by subtilisin from Bacillus subtilis may shift the procoagulant–anticoagulant equilibrium toward thrombosis

Celestine blue B as a sensor for hypochlorous acid and HOCL-modified proteins registration

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