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

Autor: Wagner Alves de Souza Judice, Graham H. Coombs, Ivarne L.S. Tersariol, Marcella Araujo Manfredi, Luiz Juliano, Tarsis F. Gesteira, Gareth D. Westrop, Sanya J. Sanderson, Cláudio S. Shida, Thiago M. Sansevero, Paulo C. Almeida, Gerson Profeta Souza
Přispěvatelé: Univ Mogi das Cruzes, Universidade Federal de São Paulo (UNIFESP), Cincinnati Childrens Hosp Med Ctr, Univ Strathclyde
Jazyk: angličtina
Rok vydání: 2013
Předmět:
Zdroj: Repositório Institucional da UNIFESP
Universidade Federal de São Paulo (UNIFESP)
instacron:UNIFESP
PLoS ONE
PLoS ONE, Vol 8, Iss 11, p e80153 (2013)
ISSN: 1932-6203
Popis: Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Cientifico Tecnologico Medical Research Council 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. Univ Mogi das Cruzes, Ctr Interdisciplinar Invest Bioquim, Mogi Das Cruzes, Brazil Universidade Federal de São Paulo, Dept Bioquim, São Paulo, Brazil Universidade Federal de São Paulo, Inst Ciencia & Tecnol, Sao Jose Dos Campos, Brazil Cincinnati Childrens Hosp Med Ctr, Div Dev Biol, Cincinnati, OH 45229 USA Universidade Federal de São Paulo, Dept Biofis, São Paulo, Brazil Univ Strathclyde, Strathclyde Inst Pharm & Biomed Sci, Glasgow, Lanark, Scotland Universidade Federal de São Paulo, Dept Bioquim, São Paulo, Brazil Universidade Federal de São Paulo, Inst Ciencia & Tecnol, Sao Jose Dos Campos, Brazil Universidade Federal de São Paulo, Dept Biofis, São Paulo, Brazil FAPESP: 2012/50219-6 Conselho Nacional de Desenvolvimento Cientifico Tecnologico: 303843/2009-8 Medical Research Council: G0700127 Web of Science
Databáze: OpenAIRE
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