Interaction of Secretory Leukocyte Protease Inhibitor with HeparinInhibits Proteases Involved in Asthma

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Fath, M.A.
Wu, X.
Hileman, R.E.
Linhardt, Robert J.
Abraham, W.M.
Kashem, M.A.
Nelson, R.M.
Wright, C.
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Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
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Protease inhibition by secretory leukocyte protease inhibitor (SLPI) is accelerated by the sulfated polysaccharides. The nature of the SLPI-polysaccharide interaction, explored with affinity chromatography, indicated that this interaction was sensitive to the charge and type of polysaccharide. Dextran and chondroitin had the lowest affinity for SLPI, followed by dermatan, heparan, and dextran sulfates. While heparin bound SLPI tightly, the highest affinity heparin chains unexpectedly contained a lower level of sulfation than more weakly interacting chains. Heparin oligosaccharides, prepared using heparin lyase I were SLPI-affinity fractionated. Surprisingly, undersulfated heparin oligosaccharides bound SLPI with the highest affinity, suggesting the importance of free hydroxyl groups for high affinity interaction. Isothermal titration calorimetry was used to determine the thermodynamics of SLPI interaction with a low molecular weight heparin, an undersulfated decasaccharide and a tetrasaccharide. The studies showed 12-14 saccharide units, corresponding to molecular weight of approximately 4,800, were required for a 1:1 (SLPI:heparin) binding stoichiometry. Furthermore, an undersulfated decasaccharide was able to bind SLPI tightly (Kd approximately 13 nM), resulting in its activation and the inhibition of neutrophil elastase and pancreatic chymotrypsin. The in vitro assessment of heparin and the decasaccharide and tetrasaccharide using stopped-flow kinetics suggested that heparin was the optimal choice to study SLPI-based in vivo protease inhibition. SLPI and heparin were co-administered by inhalation in therapy against antigen-induced airway hyperresponsiveness in a sheep bronchoprovocation model. Heparin, in combination with SLPI demonstrated in vivo efficacy reducing early and late phase bronchoconstriction. Heparin also increased the therapeutic activity of SLPI against antigen-induced airway hyperresponsiveness.
Journal of Biological Chemistry, 273, 13563-13569
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Interaction of Secretory Leukocyte Protease Inhibitor with HeparinInhibits Proteases Involved in Asthma, M.A. Fath, X. Wu, R.E. Hileman,R.J. Linhardt, W.M. Abraham, M.A. Kashem, R.M. Nelson, and C.Wright, Journal of Biological Chemistry, 273, 13563-13569, 1998.
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