Heparin modulates the endopeptidase activity of Leishmania mexicana cysteine protease cathepsin L-Like rCPB2.8

W.A.S. Judice, Marcella A Manfredi, Gerson A. Souza, Thiago M. Sansevero, Paulo C Almeida, Claudio S. Shida, Tarsis F. Gesteira, Luiz Juliano, Gareth Westrop, Sanya J Sanderson, Graham Coombs, Ivarne L.S. Tersariol

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

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. The data analysis revealed that the presence of heparin affects all steps of the enzyme reaction: (i) it decreases 3.5-fold the k1 and 4.0-fold the k−1, (ii) it affects the acyl-enzyme accumulation with pronounced decrease in k2 (2.7-fold), and also decrease in k3 (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 (Cys25)-S−/(His163)-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. 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.
LanguageEnglish
Article numbere80153
JournalPLOS One
Volume8
Issue number11
DOIs
Publication statusPublished - 21 Nov 2013

Fingerprint

Leishmania mexicana
cathepsin L
Cathepsin L
Endopeptidases
Cysteine Proteases
cysteine proteinases
heparin
Heparin
proteinases
Enzymes
glycosaminoglycans
enzymes
Glycosaminoglycans
parasites
Parasitic Diseases
glycoconjugates
Glycoconjugates
proteoglycans
Leishmania
enzymatic reactions

Keywords

  • cysteine protease
  • Leishmania mexicana
  • Heparin

Cite this

Judice, W. A. S., Manfredi, M. A., Souza, G. A., Sansevero, T. M., Almeida, P. C., Shida, C. S., ... Tersariol, I. L. S. (2013). Heparin modulates the endopeptidase activity of Leishmania mexicana cysteine protease cathepsin L-Like rCPB2.8. PLOS One, 8(11), [e80153]. https://doi.org/10.1371/journal.pone.0080153
Judice, W.A.S. ; Manfredi, Marcella A ; Souza, Gerson A. ; Sansevero, Thiago M. ; Almeida, Paulo C ; Shida, Claudio S. ; Gesteira, Tarsis F. ; Juliano, Luiz ; Westrop, Gareth ; Sanderson, Sanya J ; Coombs, Graham ; Tersariol, Ivarne L.S. / Heparin modulates the endopeptidase activity of Leishmania mexicana cysteine protease cathepsin L-Like rCPB2.8. In: PLOS One. 2013 ; Vol. 8, No. 11.
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abstract = "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. The data analysis revealed that the presence of heparin affects all steps of the enzyme reaction: (i) it decreases 3.5-fold the k1 and 4.0-fold the k−1, (ii) it affects the acyl-enzyme accumulation with pronounced decrease in k2 (2.7-fold), and also decrease in k3 (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 (Cys25)-S−/(His163)-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. 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.",
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Judice, WAS, Manfredi, MA, Souza, GA, Sansevero, TM, Almeida, PC, Shida, CS, Gesteira, TF, Juliano, L, Westrop, G, Sanderson, SJ, Coombs, G & Tersariol, ILS 2013, 'Heparin modulates the endopeptidase activity of Leishmania mexicana cysteine protease cathepsin L-Like rCPB2.8' PLOS One, vol. 8, no. 11, e80153. https://doi.org/10.1371/journal.pone.0080153

Heparin modulates the endopeptidase activity of Leishmania mexicana cysteine protease cathepsin L-Like rCPB2.8. / Judice, W.A.S.; Manfredi, Marcella A; Souza, Gerson A.; Sansevero, Thiago M.; Almeida, Paulo C; Shida, Claudio S.; Gesteira, Tarsis F.; Juliano, Luiz; Westrop, Gareth; Sanderson, Sanya J; Coombs, Graham; Tersariol, Ivarne L.S.

In: PLOS One, Vol. 8, No. 11, e80153, 21.11.2013.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Heparin modulates the endopeptidase activity of Leishmania mexicana cysteine protease cathepsin L-Like rCPB2.8

AU - Judice, W.A.S.

AU - Manfredi, Marcella A

AU - Souza, Gerson A.

AU - Sansevero, Thiago M.

AU - Almeida, Paulo C

AU - Shida, Claudio S.

AU - Gesteira, Tarsis F.

AU - Juliano, Luiz

AU - Westrop, Gareth

AU - Sanderson, Sanya J

AU - Coombs, Graham

AU - Tersariol, Ivarne L.S.

PY - 2013/11/21

Y1 - 2013/11/21

N2 - 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. The data analysis revealed that the presence of heparin affects all steps of the enzyme reaction: (i) it decreases 3.5-fold the k1 and 4.0-fold the k−1, (ii) it affects the acyl-enzyme accumulation with pronounced decrease in k2 (2.7-fold), and also decrease in k3 (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 (Cys25)-S−/(His163)-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. 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.

AB - 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. The data analysis revealed that the presence of heparin affects all steps of the enzyme reaction: (i) it decreases 3.5-fold the k1 and 4.0-fold the k−1, (ii) it affects the acyl-enzyme accumulation with pronounced decrease in k2 (2.7-fold), and also decrease in k3 (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 (Cys25)-S−/(His163)-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. 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.

KW - cysteine protease

KW - Leishmania mexicana

KW - Heparin

U2 - 10.1371/journal.pone.0080153

DO - 10.1371/journal.pone.0080153

M3 - Article

VL - 8

JO - PLOS One

T2 - PLOS One

JF - PLOS One

SN - 1932-6203

IS - 11

M1 - e80153

ER -

Judice WAS, Manfredi MA, Souza GA, Sansevero TM, Almeida PC, Shida CS et al. Heparin modulates the endopeptidase activity of Leishmania mexicana cysteine protease cathepsin L-Like rCPB2.8. PLOS One. 2013 Nov 21;8(11). e80153. https://doi.org/10.1371/journal.pone.0080153