Influence of Low Molecular Mass Heparin on the Kinetics of Neutrophil Elastase Inhibition by Mucus Proteinase Inhibitor(*) - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Journal of Biological Chemistry Année : 1995

Influence of Low Molecular Mass Heparin on the Kinetics of Neutrophil Elastase Inhibition by Mucus Proteinase Inhibitor(*)

Résumé

Commercial low molecular mass heparin accelerates the inhibition of neutrophil elastase by mucus proteinase inhibitor, the predominant antielastase of lung secretions (Faller, B., Mély, Y., Gérard, D., and Bieth, J.G. (1992) Biochemistry 31, 8285-8290). To study the kinetic mechanism of this rate enhancement, we have isolated a 4.5-kDa heparin fragment from commercial heparin. This compound is fairly monodisperse as shown by analytical ultracentrifugation. It binds elastase and inhibitor with a 1:1 stoichiometry and an equilibrium dissociation constant of 3 and 210 nM, respectively. It also forms a tight complex with EI. Flow calorimetry shows that the inhibitor-heparin interaction is characterized by a large negative enthalpy change (delta H0 = -45.2 kJ mol-1) and a small entropy change (delta S = -23.7 J K-1 mol-1). Stopped-flow kinetics run under pseudo-first-order conditions ([Io] > [Eo]) show that in the absence of heparin the inhibition conforms to a simple bimolecular reaction, [formula: see text] where, kass = 3.1 x 10(6) M-1 s-1, kdiss = 10(-4) s-1, and Ki = 33 pM, whereas in the presence of heparin, E and I react via a two-step mechanism, [formula: see text] where Ki* = 86 nM, k2 = 2.2 s-1, k-2 = 10(-3) s-1, and Ki = 37 pM. Thus, heparin increases both the rate of inhibition by promoting the formation of a high affinity EI* intermediate and the rate of EI dissociation. Since the dissociation is negligible in bronchial secretions where the inhibitor concentration is much higher than Ki, it may be concluded that heparin significantly potentiates the inhibitor's antielastase potential in vivo.

Dates et versions

hal-02150493 , version 1 (07-06-2019)

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Citer

Martine Cadène, Christian Boudier, Guy Daney de Marcillac, Joseph Bieth. Influence of Low Molecular Mass Heparin on the Kinetics of Neutrophil Elastase Inhibition by Mucus Proteinase Inhibitor(*). Journal of Biological Chemistry, 1995, 270 (22), pp.13204-13209. ⟨10.1074/jbc.270.22.13204⟩. ⟨hal-02150493⟩

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