Inhibition of leucocyte elastase by heparin and its derivatives

Rédini, Françoise; Tixier, J. M.; Petitou, Maurice; Choay, J; Robert, L.; Hornebeck, William

doi:10.1042/bj2520515

Cited by 132 publications

(86 citation statements)

References 26 publications

(34 reference statements)

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“…Sulfated glycosaminoglycans prevent NE-induced lung emphysema in animal models (31,33). These ligands are partial inhibitors of the elastolytic activity of NE (27,34,35). Suramin, which fully inhibits elastolysis and has already been used in human cancer therapy (5)(6)(7)(8) should therefore prove to be an efficient drug in diseases characterized by massive neutrophil recruitment and activation.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of Neutrophil Serine Proteinases by Suramin

Cadène

Duranton

North

et al. 1997

Journal of Biological Chemistry

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Suramin, a hexasulfonated naphtylurea recently used as an anti-tumor drug, is a potent inhibitor of human neutrophil elastase, cathepsin G, and proteinase 3. The complexes it forms with these enzymes are partially active on synthetic substrates, but full inhibition takes place when elastase activity is measured with fibrous elastin or when cathepsin G activity is measured using platelet aggregation. One molecule of elastase binds four molecules of suramin with a K i of 2 ؋ 10 ؊7 M as determined by enzyme inhibition or intrinsic fluorescence enhancement of suramin. The binding curves show no sign of cooperativity or anticooperativity. The K i for the complexes with cathepsin G and proteinase 3 are 8 ؋ 10 ؊8 and 5 ؋ 10 ؊7 M, respectively. Ionic strength increases the K i of the elastase-suramin complex in a way that suggests that four of the six sulfonate groups of suramin form ionic interactions with basic residues of the enzyme and that at saturation almost all arginines of elastase form salt bridges with suramin. The neutrophil proteinase-inhibitory activity of suramin might be used to prevent tissue destruction and thrombus formation in diseases where massive infiltration and activation of neutrophils take place.

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Section: Discussionmentioning

confidence: 99%

Inhibition of Neutrophil Serine Proteinases by Suramin

Cadène

Duranton

North

et al. 1997

Journal of Biological Chemistry

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“…The effect of DNA on the NE/MPI system is reminiscent of that observed with heparin, another anionic polymer. Heparin is also a hyperbolic inhibitor of NE [13] which forms a tight complex with MPI [11]. However, unlike DNA, heparin strongly potentiates the inhibition of NE by MPI [11].…”

Section: Discussionmentioning

confidence: 99%

DNA binds neutrophil elastase and mucus proteinase inhibitor and impairs their functional activity

Belorgey

Bieth

1995

FEBS Letters

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DNA binds neutrophil elastase and mucus proteinase inhibitor as evidenced by affinity chromatography on elastaseSepharose, inhibitor-Sepharose and DNA-cellulose. DNA is a potent hyperbolic inhibitor of elastase. The polynucleotide-en~yme complex is partially active on synthetic substrates and on elastin. DNA strongly increases kd~ and Ki for the inhibition of elastase by mucus proteinase inhibitor kass t E + I ~ EI, K i = kdi~Jka~s).kdiss ['he above effects are all salt-dependent. At physiological ionic strength, DNA is a potent inhibitor of the elastolytic activity of elastase and increases k~ and K~ for the elastase-mucus proteinase inhibitor interaction 160-fold and 100-fold, respectively.,~ey words': Elastase; Mucus proteinase inhibitor; Enzyme i:inetics; DNA; Cystic fibrosis L. IntroductionNeutrophil elastase (NE) is a 30-kDa cationic glycoprotein .vhose crystal structure is known. This serine proteinase cleaves t number of plasma and extracellular matrix proteins including ,.:lastin. It is stored in the azurophil granules of neutrophils from ,vhich it may be released following excessive phagocytosis or :ell death. Its extracellular proteolytic action is normally pre-,lented by protein proteinase inhibitors such as ~l-proteinase nhibitor, ~2-macroglobulin or mucus proteinase inhibitor MPI) (for a review see [1]). The latter is an 11.7-kDa unglyco~ylated cationic protein composed of two domains of similar ;ize and architecture ([2] and refs. therein).MPI is the most abundant NE inhibitor of airways secretions ~here it occurs in concentrations as high as 5/IM [3]. Despite ~:his high load of inhibitor, bronchial secretions from patients ~vith cystic fibrosis contain significant amounts of active NE [4]. On the other hand, these secretions also contain large quantities ~f neutrophil-derived DNA [5]. This raises the question as to uhether DNA interferes with the inhibition of NE by MPI. The ~resent investigation attempts to answer this question. Materials and methods MaterialsHuman NE was purified from purulent sputum as described in [6]. Recombinant MPI was obtained from Synergen, Boulder, CO through the courtesy of Dr. H.P. Schnebli, Ciba-Geigy, Basel, Switzerland. NE and MPI were active site titrated as indicated in [6]. Stock solutions of Suc-Ala3-pNA and MeOSuc-Ala2-Pro-Val-pNA (Bachem, Bubendorf, Switzerland) were made in N-methylpyrolidone and dimethylformamide, respectively. The final concentration of organic solvent in the reaction mixtures was 2% (v/v) throughout. Remazol Brilliant-blue elastin (RBB-elastin) was purchased from Elastin Products Company (Owenville, MO). Salmon sperm DNA and DNA~cellulose came from Sigma. NE or MPI were coupled to epoxy-activated Sepharose (Pharmacia) as described by the manufacturer. The three affinity supports (4 ml) were poured into HR 10/10 Pharmacia columns and equilibrated with 50 mM HEPES buffer, pH 7.4. Determination of k,,, and K,~The kinetic constants kca t and Km for the hydrolysis of MeOSuc-Ala2-Pro-Val-pNA by 30 nM free or DNA-bound NE were determine...

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“…However, a possible immunomodulatory function is suggested, as increasing evidence, both experimental and clinical, indicates heparin to possess a range of anti-in¯ammatory properties (reviewed by Jaques, 1979;Tyrrell et al, 1999). Many of these e ects may be mediated through the ability of this large, negatively charged molecule to bind and inactivate an array of in¯ammatory proteins, including certain complement components (Matzner et al, 1984;Strunk & Colten, 1976), chemokines (Miller & Krangel, 1992) and products of activated granulocytes (Fredens et al, 1991;Redini et al, 1988;Walsh et al, 1991b) as well as thrombin, a known proin¯ammatory mediator which acts through PAR-1 receptor activation. In addition, heparin is known to bind a number of adhesion molecules involved in leucocyte tra cking into tissues, including mac-1 (CD11b/CD18; Diamond et al, 1995) and L-selectin (Koenig et al, 1998) on in¯ammatory cells and the endothelial adhesion molecules P-selectin (Revelle et al, 1996;Skinner et al, 1991) and platelet endothelial adhesion molecule-1 (PECAM-1; Watt et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

The effects of heparin on the adhesion of human peripheral blood mononuclear cells to human stimulated umbilical vein endothelial cells

Smailbegovic

Lever

Page

2001

British J Pharmacology

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1 The e ects of unfractionated heparin (UH) and a selectively O-desulphated derivative of heparin (ODSH), lacking anticoagulant activity, on the adhesion of human peripheral blood mononuclear cells (HPBMNC) to human stimulated umbilical vein endothelial cells (HUVECs), were investigated. 2 For comparison, the e ects of poly-L-glutamic acid (PGA), a large polyanionic molecule without sulphate groups and two di erent molecular weight sulphated dextrans (DS 5 k and DS 10 k) were studied. 3 UH (50 ± 1000 u ml 71 ) signi®cantly (P50.05) inhibited the adhesion of HPBMNC to HUVECs, stimulated with IL-1b (100 u ml 71 ), TNF-a (1000 u ml 71 ) or LPS (100 mg ml 71 ), when the drugs were added together with stimuli to HUVECs and coincubated for 6 h. Such e ects on adhesion occurred with limited in¯uence on expression of relevant endothelial adhesion molecules (ICAM-1 and VCAM-1). 4 UH (100 ± 1000 u ml 71 ), when added to prestimulated HUVECs, signi®cantly (P50.05) increased adhesion of mononuclear cells to endothelium at the higher concentrations tested, without any e ect on adhesion molecule expression. In contrast, the opposite e ect was observed when human polymorphonuclear leucocyte adhesion was examined, under the same experimental conditions, suggesting that the observed potentiation of HPBMNC adhesion is cell speci®c. 5 The e ects of UH on HPBMNC adhesion were shared by the non-anticoagulant ODSH (600 ± 6000 mg ml 71 ) but not by sulphated dextrans or PGA (300 ± 6000 mg ml 71 ). 6 Heparin a ects the adhesion of HPBMNC to stimulated endothelium, in both an inhibitory and potentiating manner, e ects which are unrelated to its anticoagulant activity and not solely dependent on molecular charge characteristics.

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Inhibition of leucocyte elastase by heparin and its derivatives

Cited by 132 publications

References 26 publications

Inhibition of Neutrophil Serine Proteinases by Suramin

Inhibition of Neutrophil Serine Proteinases by Suramin

DNA binds neutrophil elastase and mucus proteinase inhibitor and impairs their functional activity

The effects of heparin on the adhesion of human peripheral blood mononuclear cells to human stimulated umbilical vein endothelial cells

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