Heparin modulates the endopeptidase activity of Leishmania mexicana cysteine protease cathepsin L-Like rCPB2.8.
<h4>Background</h4>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 inte...
Saved in:
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2013-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0080153 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849331787361157120 |
|---|---|
| author | Wagner A S Judice Marcella A Manfredi Gerson P Souza Thiago M Sansevero Paulo C Almeida Cláudio S Shida Tarsis F Gesteira Luiz Juliano Gareth D Westrop Sanya J Sanderson Graham H Coombs Ivarne L S Tersariol |
| author_facet | Wagner A S Judice Marcella A Manfredi Gerson P Souza Thiago M Sansevero Paulo C Almeida Cláudio S Shida Tarsis F Gesteira Luiz Juliano Gareth D Westrop Sanya J Sanderson Graham H Coombs Ivarne L S Tersariol |
| author_sort | Wagner A S Judice |
| collection | DOAJ |
| description | <h4>Background</h4>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.<h4>Methodology/principal findings</h4>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.<h4>Conclusions/significance</h4>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 | Article |
| id | doaj-art-0147605441e94b73a97ea1a6f9f4b77a |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-0147605441e94b73a97ea1a6f9f4b77a2025-08-20T03:46:24ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-01811e8015310.1371/journal.pone.0080153Heparin modulates the endopeptidase activity of Leishmania mexicana cysteine protease cathepsin L-Like rCPB2.8.Wagner A S JudiceMarcella A ManfrediGerson P SouzaThiago M SanseveroPaulo C AlmeidaCláudio S ShidaTarsis F GesteiraLuiz JulianoGareth D WestropSanya J SandersonGraham H CoombsIvarne L S Tersariol<h4>Background</h4>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.<h4>Methodology/principal findings</h4>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.<h4>Conclusions/significance</h4>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.https://doi.org/10.1371/journal.pone.0080153 |
| spellingShingle | Wagner A S Judice Marcella A Manfredi Gerson P Souza Thiago M Sansevero Paulo C Almeida Cláudio S Shida Tarsis F Gesteira Luiz Juliano Gareth D Westrop Sanya J Sanderson Graham H Coombs Ivarne L S Tersariol Heparin modulates the endopeptidase activity of Leishmania mexicana cysteine protease cathepsin L-Like rCPB2.8. PLoS ONE |
| 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 |
| url | https://doi.org/10.1371/journal.pone.0080153 |
| work_keys_str_mv | AT wagnerasjudice heparinmodulatestheendopeptidaseactivityofleishmaniamexicanacysteineproteasecathepsinllikercpb28 AT marcellaamanfredi heparinmodulatestheendopeptidaseactivityofleishmaniamexicanacysteineproteasecathepsinllikercpb28 AT gersonpsouza heparinmodulatestheendopeptidaseactivityofleishmaniamexicanacysteineproteasecathepsinllikercpb28 AT thiagomsansevero heparinmodulatestheendopeptidaseactivityofleishmaniamexicanacysteineproteasecathepsinllikercpb28 AT paulocalmeida heparinmodulatestheendopeptidaseactivityofleishmaniamexicanacysteineproteasecathepsinllikercpb28 AT claudiosshida heparinmodulatestheendopeptidaseactivityofleishmaniamexicanacysteineproteasecathepsinllikercpb28 AT tarsisfgesteira heparinmodulatestheendopeptidaseactivityofleishmaniamexicanacysteineproteasecathepsinllikercpb28 AT luizjuliano heparinmodulatestheendopeptidaseactivityofleishmaniamexicanacysteineproteasecathepsinllikercpb28 AT garethdwestrop heparinmodulatestheendopeptidaseactivityofleishmaniamexicanacysteineproteasecathepsinllikercpb28 AT sanyajsanderson heparinmodulatestheendopeptidaseactivityofleishmaniamexicanacysteineproteasecathepsinllikercpb28 AT grahamhcoombs heparinmodulatestheendopeptidaseactivityofleishmaniamexicanacysteineproteasecathepsinllikercpb28 AT ivarnelstersariol heparinmodulatestheendopeptidaseactivityofleishmaniamexicanacysteineproteasecathepsinllikercpb28 |