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Papers In Press, published online ahead of print October 2, 2000
Centro Interdisciplinar de Investigacoes Bioquimica, Universidade de Mogi das Cruzes, Mogi das Cruzes, Sao Paulo 08780-911
Corresponding Author: ivarne{at}ccb.umc.br
ABSTRACT It has been shown that lysosomal cysteine proteinases, specially cathepsin B, has been implicated in a variety of diseases involving tissue remodeling states, such as inflammation, parasite infection, and tumor metastasis, by degradation of extracellular matrix components. Recently, we have shown that heparin and heparan sulfate bind to papain specifically , this interaction induces an increase of its a(alpha)-helix content and stabilizes the enzyme structure even at alkaline pH (Almeida, P. C., Nantes, I. L., Rizzi, C. C. A., Júdice, W. A. S., Chagas, J. R., Juliano, L., Nader, H. B., and Tersariol, I. L. S. (1999) J. Biol. Chem. 274, 30433-30438). In the present work, a combination of circular dichroism analysis, affinity chromatography, cathepsin B mutants, and fluorogenic substrate assays were used to characterize the interaction of human cathepsin B with glycosaminoglycans. The nature of the cathepsin B-glycosaminoglycans interaction was sensitive to the charge and type of polysaccharide. Like papain, heparin and heparan sulfate bind cathepsin B specifically, and this interaction reduces the loss of cathepsin B a(alpha)-helix content at alkaline pH. Our data show that the coupling of cathepsin B with heparin or heparan sulfate can potentiate the endopeptidase activity of the cathepsin B, increasing 5-fold the half-life (t1/2) of the enzyme at alkaline pH. Most of these effects are related to the interaction of heparin and heparan sulfate with His111 residue of the cathepsin B occluding loop. These results strongly suggest that heparan sulfate may be an important binding site for cathepsin B at cell surface, reporting a novel physiological role for heparan sulfate proteoglycans.
J. Biol. Chem, 10.1074/jbc.M003820200
Submitted on May 4, 2000
Revised on September 11, 2000
Accepted on September 29, 2000
Cathepsin B activity regulation heparin-like glycosaminoglycans protect human cathepsin B from alkaline PH-induced inactivation
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