1 H, 15 N and 13C assignments of the cysteine protease inhibitor Chagasin from Trypanosoma cruzi

2006 ◽  
Vol 36 (S1) ◽  
pp. 30-30
Author(s):  
Rodolpho do Aido-Machado ◽  
Didier Salmon ◽  
Anne Diehl ◽  
Martina Leidert ◽  
Oliver Schmetzer ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Protease Inhibitor ◽  
Cysteine Protease ◽  
Cysteine Protease Inhibitor

scholarly journals Tigutcystatin, a cysteine protease inhibitor from Triatoma infestans midgut expressed in response to Trypanosoma cruzi

2011 ◽  
Vol 413 (2) ◽  
pp. 241-247 ◽  
Author(s):  
Diego S. Buarque ◽  
Letícia M.N. Spindola ◽  
Rafael M. Martins ◽  
Glória R.C. Braz ◽  
Aparecida S. Tanaka
Keyword(s):  
Trypanosoma Cruzi ◽  
Protease Inhibitor ◽  
Cysteine Protease ◽  
Cysteine Protease Inhibitor ◽  
Triatoma Infestans

Crystal structure of chagasin, the endogenous cysteine-protease inhibitor from Trypanosoma cruzi

2007 ◽  
Vol 157 (2) ◽  
pp. 416-423 ◽  
Author(s):  
Aline Almeida Figueiredo da Silva ◽  
Leandro de Carvalho Vieira ◽  
Marco Aurélio Krieger ◽  
Samuel Goldenberg ◽  
Nilson Ivo Tonin Zanchin ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Trypanosoma Cruzi ◽  
Protease Inhibitor ◽  
Cysteine Protease ◽  
Cysteine Protease Inhibitor

Solution Structure and Backbone Dynamics of the Trypanosoma cruzi Cysteine Protease Inhibitor Chagasin

2006 ◽  
Vol 357 (5) ◽  
pp. 1511-1521 ◽  
Author(s):  
Didier Salmon ◽  
Rodolpho do Aido-Machado ◽  
Anne Diehl ◽  
Martina Leidert ◽  
Oliver Schmetzer ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Protease Inhibitor ◽  
Cysteine Protease ◽  
Solution Structure ◽  
Backbone Dynamics ◽  
Cysteine Protease Inhibitor

scholarly journals A Cysteine Protease Inhibitor Protects Dogs from Cardiac Damage during Infection by Trypanosoma cruzi

2005 ◽  
Vol 49 (12) ◽  
pp. 5160-5161 ◽  
Author(s):  
S. C. Barr ◽  
K. L. Warner ◽  
B. G. Kornreic ◽  
J. Piscitelli ◽  
A. Wolfe ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Protease Inhibitor ◽  
Chagas Disease ◽  
Cysteine Protease ◽  
Therapeutic Target ◽  
Troponin I ◽  
Myocardial Damage ◽  
Cysteine Protease Inhibitor ◽  
Cardiac Damage ◽  
Histopathological Lesion

ABSTRACT Cruzain is an essential cysteine protease of Trypanosoma cruzi and a therapeutic target for Chagas' disease. Eight dogs were infected with T. cruzi; three were treated with an inhibitor of cruzain, K777, for 14 days. Treatment with K777 abrogated myocardial damage by T. cruzi, as documented by histopathological lesion scores and serum troponin I levels.

Upregulation of the secretory pathway in cysteine protease inhibitor-resistant Trypanosoma cruzi

2000 ◽  
Vol 113 (8) ◽  
pp. 1345-1354
Author(s):  
J.C. Engel ◽  
C.T. Garcia ◽  
I. Hsieh ◽  
P.S. Doyle ◽  
J.H. McKerrow
Keyword(s):  
Trypanosoma Cruzi ◽  
Protease Inhibitor ◽  
Golgi Complex ◽  
Cysteine Protease ◽  
Secretory Pathway ◽  
Complex Function ◽  
Cysteine Protease Inhibitor ◽  
Cross Resistance ◽  
Structural Levels

A novel chemotherapy in development for Chagas' disease targets cruzain, the major cysteine protease of Trypanosoma cruzi. Peptidomimetic inhibitors disrupt the intracellular cycle of the parasite and rescue animals from a lethal infection. Inhibitor killing of parasites results from interruption of autocatalytic cruzain processing and transport to lysosomes, and massive accumulation of precursor protein in the Golgi complex. To further understand the mechanisms of protease processing and transport in this primitive eukaryote, and uncover potential mechanisms for resistance to these drugs, we generated cysteine-protease inhibitor (CPI)-resistant epimastigotes in vitro and investigated the mechanisms involved at the biochemical and structural levels. Resistance to 20-fold the lethal CPI concentration, achieved after a year of gradual drug increase, was accompanied by a modest decrease in growth rate. A marked increase in the number of vesicles trafficking from the Golgi complex to the flagellar pocket occurs in resistant cells. No mature protease reaches lysosomes though accumulation of endocytosed gold particles in lysosomes appears to be normal. Higher molecular mass cruzain species, consistent with complexes of cruzain precursors and inhibitor, are secreted by CPI-resistant parasites into the culture supernatant. Release of these cruzain precursors may be facilitated by an enhanced acidification of trans-Golgi cisternae in resistant parasites. The pH within Golgi cisternae is higher in control epimastigotes and most mature cruzain is lysosomal. Cruzain activity is negligible in CPI-resistant epimastigote extracts compared to the parental clone. Activity is restored following withdrawal of the inhibitor. No cross-resistance to the therapeutic drugs nifurtimox and benznidazole occurred and, conversely, parasites resistant to these drugs were sensitive to CPI. Protease inhibitors are thus potential therapeutical alternatives in cases of nifurtimox/benznidazole resistance. Cumulatively, these results suggest that CPI-resistance induces upregulation of Golgi complex function and post-Golgi secretory pathway, and release of precursors before the enzyme reaches its site of biologic activity.

Cysteine protease inhibitors alter Golgi complex ultrastructure and function in Trypanosoma cruzi

1998 ◽  
Vol 111 (5) ◽  
pp. 597-606 ◽  
Author(s):  
J.C. Engel ◽  
P.S. Doyle ◽  
J. Palmer ◽  
I. Hsieh ◽  
D.F. Bainton ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Protease Inhibitor ◽  
Protease Inhibitors ◽  
Golgi Complex ◽  
Cysteine Protease ◽  
Intracellular Localization ◽  
Fluorescent Microscopy ◽  
Brefeldin A ◽  
Cysteine Protease Inhibitor ◽  
Cysteine Protease Inhibitors

Cruzain, the major cysteine protease of the protozoan parasite Trypanosoma cruzi, is a target of rational drug design for chemotherapy of Chagas' disease. The precise biological role of cruzain in the parasite life cycle and the mechanism involved in the trypanocidal effect of cysteine protease inhibitors are still unclear. Here we report biological and ultrastructural alterations caused by cysteine protease inhibitors in T. cruzi epimastigotes. Cruzain, a glycoprotein that transits the Golgi-endosomal pathway, localized to pre-lysosomes/lysosomes in the posterior end of untreated epimastigotes by fluorescent microscopy utilizing either a biotinylated cysteine protease inhibitor to tag the active site, or a specific anti-cruzain antibody. Radiolabeled or biotinylated cysteine protease inhibitors bound exclusively to cruzain in intact epimastigotes confirming that cruzain is accessible to, and is targeted by the inhibitors. Treatment of T. cruzi epimastigotes with specific cysteine protease inhibitors arrested growth, altered the intracellular localization of cruzain, and induced major alterations in the Golgi complex. Following treatment, cruzain accumulated in peripheral dilations of Golgi cisternae. There was a concomitant 70% reduction in gold-labeled cruzain transported to lysosomes. Cisternae abnormalities in the Golgi compartment were followed by distention of ER and nuclear membranes. Brefeldin A increased the number and size of cisternae in epimastigotes. Pre-treatment of epimastigotes with cysteine protease inhibitors followed by exposure to brefeldin A induced a more rapid appearance of the cysteine protease inhibitor-induced Golgi alterations. Our results suggest that cysteine protease inhibitors prevent the normal autocatalytic processing and trafficking of cruzain within the Golgi apparatus. Accumulation of cruzain may decrease mobility of Golgi membranes and result in peripheral distention of cisternae. These major alterations of the Golgi complex parallel the death of T. cruzi epimastigotes.

scholarly journals A New Cruzipain-Mediated Pathway of Human Cell Invasion by Trypanosoma cruzi Requires Trypomastigote Membranes

2004 ◽  
Vol 72 (10) ◽  
pp. 5892-5902 ◽  
Author(s):  
Isabela M. Aparicio ◽  
Julio Scharfstein ◽  
Ana Paula C. A. Lima
Keyword(s):  
Trypanosoma Cruzi ◽  
Protease Inhibitor ◽  
Host Cell ◽  
Cell Invasion ◽  
Cysteine Protease ◽  
Mammalian Cells ◽  
Host Cells ◽  
Cysteine Protease Inhibitor ◽  
Host Cell Invasion ◽  
Different Strains

ABSTRACT The intracellular protozoan Trypanosoma cruzi causes Chagas' disease, a chronic illness associated with cardiomyopathy and digestive disorders. This pathogen invades mammalian cells by signaling them through multiple transduction pathways. We previously showed that cruzipain, the main cysteine protease of T. cruzi, promotes host cell invasion by activating kinin receptors. Here, we report a cruzipain-mediated invasion route that is not blocked by kinin receptor antagonists. By testing different strains of T. cruzi, we observed a correlation between the level of cruzipain secreted by trypomastigotes and the capacity of the pathogen to invade host cells. Consistent with a role for cruzipain, the cysteine protease inhibitor N-methylpiperazine-urea-Phe-homophenylalanine-vinylsulfone-benzene impaired the invasion of human smooth muscle cells by strains Dm28c and X10/6 but not by the G isolate. Cruzipain-rich supernatants of Dm28c trypomastigotes enhanced the infectivity of isolate G parasites twofold, an effect which was abolished by the cysteine protease inhibitor l-trans-epoxysuccinyl-leucylamido-(4-guanidino)butane and by thapsigargin, a drug that induces depletion of the intracellular Ca2+ stores. The enhancement due to Dm28 supernatants was abolished upon cruzipain immunodepletion, and the activity was restored by purified cruzipain. In contrast, supernatants from isolate G trypomastigotes (with low levels of cruzipain) or supernatants from Dm28c epimastigotes or purified cruzipain alone did not enhance parasite invasion, indicating that the protease is required but not sufficient to engage this invasion pathway. We provide evidence that activation of this pathway requires cruzipain-mediated processing of a trypomastigote molecule associated with parasite-shed membranes. Our results couple cruzipain to host cell invasion through a kinin-independent route and further suggest that high-level cruzipain expression may contribute to parasite infectivity.

scholarly journals A Novel Cysteine Protease Inhibitor of Naegleria fowleri That Is Specifically Expressed during Encystation and at Mature Cysts

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 388
Author(s):  
Hương Giang Lê ◽  
A-Jeong Ham ◽  
Jung-Mi Kang ◽  
Tuấn Cường Võ ◽  
Haung Naw ◽  
...  
Keyword(s):  
Protease Inhibitor ◽  
Cysteine Protease ◽  
Expression Profiles ◽  
Critical Role ◽  
Cyst Formation ◽  
Cysteine Proteases ◽  
Cysteine Protease Inhibitor ◽  
Naegleria Fowleri ◽  
Nægleria Fowleri ◽  
Cystatin Family

Naegleria fowleri is a free-living amoeba that is ubiquitous in diverse natural environments. It causes a fatal brain infection in humans known as primary amoebic meningoencephalitis. Despite the medical importance of the parasitic disease, there is a great lack of knowledge about the biology and pathogenicity of N. fowleri. In this study, we identified and characterized a novel cysteine protease inhibitor of N. fowleri (NfCPI). NfCPI is a typical cysteine protease inhibitor belonging to the cystatin family with a Gln-Val-Val-Ala-Gly (QVVAG) motif, a characteristic motif conserved in the cystatin family of proteins. Bacterially expressed recombinant NfCPI has a dimeric structure and exhibits inhibitory activity against several cysteine proteases including cathespin Bs of N. fowleri at a broad range of pH values. Expression profiles of nfcpi revealed that the gene was highly expressed during encystation and cyst of the amoeba. Western blot and immunofluorescence assays also support its high level of expression in cysts. These findings collectively suggest that NfCPI may play a critical role in encystation or cyst formation of N. fowleri by regulating cysteine proteases that may mediate encystation or mature cyst formation of the amoeba. More comprehensive studies to investigate the roles of NfCPI in encystation and its target proteases are necessary to elucidate the regulatory mechanism and the biological significance of NfCPI.

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