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Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8
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,...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3836952/ https://www.ncbi.nlm.nih.gov/pubmed/24278253 http://dx.doi.org/10.1371/journal.pone.0080153 |
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author | Judice, Wagner A. S. Manfredi, Marcella A. Souza, Gerson P. Sansevero, Thiago M. Almeida, Paulo C. Shida, Cláudio S. Gesteira, Tarsis F. Juliano, Luiz Westrop, Gareth D. Sanderson, Sanya J. Coombs, Graham H. Tersariol, Ivarne L. S. |
author_facet | Judice, Wagner A. S. Manfredi, Marcella A. Souza, Gerson P. Sansevero, Thiago M. Almeida, Paulo C. Shida, Cláudio S. Gesteira, Tarsis F. Juliano, Luiz Westrop, Gareth D. Sanderson, Sanya J. Coombs, Graham H. Tersariol, Ivarne L. S. |
author_sort | Judice, Wagner A. S. |
collection | PubMed |
description | 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 ΔG = 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 α-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. |
format | Online Article Text |
id | pubmed-3836952 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-38369522013-11-25 Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 Judice, Wagner A. S. Manfredi, Marcella A. Souza, Gerson P. Sansevero, Thiago M. Almeida, Paulo C. Shida, Cláudio S. Gesteira, Tarsis F. Juliano, Luiz Westrop, Gareth D. Sanderson, Sanya J. Coombs, Graham H. Tersariol, Ivarne L. S. PLoS One Research Article 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 ΔG = 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 α-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. Public Library of Science 2013-11-21 /pmc/articles/PMC3836952/ /pubmed/24278253 http://dx.doi.org/10.1371/journal.pone.0080153 Text en © 2013 Tersariol et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Judice, Wagner A. S. Manfredi, Marcella A. Souza, Gerson P. Sansevero, Thiago M. Almeida, Paulo C. Shida, Cláudio S. Gesteira, Tarsis F. Juliano, Luiz Westrop, Gareth D. Sanderson, Sanya J. Coombs, Graham H. Tersariol, Ivarne L. S. Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
title | Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
title_full | Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
title_fullStr | Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
title_full_unstemmed | Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
title_short | Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
title_sort | heparin modulates the endopeptidase activity of leishmania mexicana cysteine protease cathepsin l-like rcpb2.8 |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3836952/ https://www.ncbi.nlm.nih.gov/pubmed/24278253 http://dx.doi.org/10.1371/journal.pone.0080153 |
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