Author |
Judice, Wagner A. S.
![]() Manfredi, Marcella A. ![]() Souza, Gerson P. ![]() Sansevero, Thiago M. ![]() Almeida, Paulo C. ![]() ![]() Shida, Claudio S. ![]() ![]() Gesteira, Tarsis F. ![]() Juliano, Luiz ![]() ![]() Westrop, Gareth D. ![]() Sanderson, Sanya J. ![]() Coombs, Graham H. ![]() Tersariol, Ivarne Luis dos Santos ![]() ![]() |
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 |
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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 |
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Access rights | Open access ![]() |
Type | Article |
Web of Science ID | WOS:000327539800059 |
URI | http://repositorio.unifesp.br/handle/11600/36988 |
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