Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8

Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8

Author Judice, Wagner A. S. Google Scholar
Manfredi, Marcella A. Google Scholar
Souza, Gerson P. Google Scholar
Sansevero, Thiago M. Google Scholar
Almeida, Paulo C. Autor UNIFESP Google Scholar
Shida, Claudio S. Autor UNIFESP Google Scholar
Gesteira, Tarsis F. Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Westrop, Gareth D. Google Scholar
Sanderson, Sanya J. Google Scholar
Coombs, Graham H. Google Scholar
Tersariol, Ivarne Luis dos Santos Autor UNIFESP Google Scholar
Institution Univ Mogi das Cruzes
Universidade Federal de São Paulo (UNIFESP)
Cincinnati Childrens Hosp Med Ctr
Univ Strathclyde
Abstract Background: Cysteine protease B is considered crucial for the survival and infectivity of the Leishmania in its human host. Several microorganism pathogens bind to the heparin-like glycosaminoglycans chains of proteoglycans at host-cell surface to promote their attachment and internalization. Here, we have investigated the influence of heparin upon Leishmania mexicana cysteine protease rCPB2.8 activity.Methodology/Principal Findings: the data analysis revealed that the presence of heparin affects all steps of the enzyme reaction: (i) it decreases 3.5-fold the k(1) and 4.0-fold the k(-1), (ii) it affects the acyl-enzyme accumulation with pronounced decrease in k(2) (2.7-fold), and also decrease in k(3) (3.5-fold). the large values of triangle G = 12 kJ/mol for the association and dissociation steps indicate substantial structural strains linked to the formation/dissociation of the ES complex in the presence of heparin, which underscore a conformational change that prevents the diffusion of substrate in the rCPB2.8 active site. Binding to heparin also significantly decreases the alpha-helix content of the rCPB2.8 and perturbs the intrinsic fluorescence emission of the enzyme. the data strongly suggest that heparin is altering the ionization of catalytic (Cys(25))-S-/(His(163))-Im(+) H ion pair of the rCPB2.8. Moreover, the interaction of heparin with the N-terminal pro-region of rCPB2.8 significantly decreased its inhibitory activity against the mature enzyme.Conclusions/Significance: Taken together, depending on their concentration, heparin-like glycosaminoglycans can either stimulate or antagonize the activity of cysteine protease B enzymes during parasite infection, suggesting that this glycoconjugate can anchor parasite cysteine protease at host cell surface.
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Cientifico Tecnologico
Medical Research Council
Grant number FAPESP: 2012/50219-6
Conselho Nacional de Desenvolvimento Cientifico Tecnologico: 303843/2009-8
Medical Research Council: G0700127
Date 2013-11-21
Published in Plos One. San Francisco: Public Library Science, v. 8, n. 11, 12 p., 2013.
ISSN 1932-6203 (Sherpa/Romeo, impact factor)
Publisher Public Library Science
Extent 12
Origin http://dx.doi.org/10.1371/journal.pone.0080153
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000327539800059
URI http://repositorio.unifesp.br/handle/11600/36988

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