Expression and cellular localisation of Trypanosoma cruzi calpains

Ennes-Vidal, Vítor; Pitaluga, André Nóbrega; Britto, Constança; Branquinha, Marta H.; Santos, André Luis Souza dos; Menna-Barreto, Rubem Figueiredo Sadok; d’Avila-Levy, Claudia M.

doi:10.1590/0074-02760200142

Cited by 3 publications

(11 citation statements)

References 31 publications

(79 reference statements)

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“…Moreover, the classification of calpains in families is constantly being updated. In general, non-mammalian organisms have few copies of these non-classical calpains in their genomes, but in trypanosomatids, a massive expansion of calpain genes took place, sometimes harboring additional domains with the most varied functions and origins [ 8 , 10 , 11 , 29 ].…”

Section: Calpain Superfamily Expansion In Trypanosomatid Genomes Amentioning

confidence: 99%

“…The authors found 53 T. cruzi calpain sequences, eight of them carrying the 204 bp repeats of H49 proteins associated with the calpain-related sequences, which were referred to as H49/calpains. Then, our research group screened the genome of the T. cruzi CL Brener strain to identify and classify the calpain sequences, domain arrangements, and gene expression patterns among epimastigotes, amastigotes, and trypomastigotes [ 11 ]. Our analysis disclosed 63 calpain sequences with 14 different domain arrangements.…”

Section: Calpain Superfamily Expansion In Trypanosomatid Genomes Amentioning

confidence: 99%

“…The comparison of calpain mRNA abundance by qPCR revealed: (i) one highly expressed only in amastigotes (ii) three significantly upmodulated in amastigotes but downregulated in trypomastigotes; (iii) one upmodulated only in trypomastigotes; (iv) five significantly upmodulated in the insect form in comparison to the clinically relevant forms; (v) two without modulation between epimastigotes and amastigotes, but downregulated in trypomastigotes; and (vi) two constitutive expression sequences. Probably, this surprising expansion of calpain sequences and their differential expression between the life-cycle forms of the parasite may be correlated to distinct functions, such as the need for survival or growth within several distinctive environments, like the mammalian host and the insect vector [ 5 , 8 , 11 ]. An overview of calpain gene expression is summarized in Table 2 .…”

Section: Calpain Superfamily Expansion In Trypanosomatid Genomes Amentioning

confidence: 99%

“…Fortunately, the genome of A. deanei was recently sequenced and assembled [ 42 ], making feasible a screening for calpain sequences in this organism, allowing the identification of genes that keep the basic requisites to retain proteolytic activity. Therefore, here, we screened the A. deanei genome using previously described approaches [ 10 , 11 ]. Our search revealed the presence of 65 calpain sequences distributed in two of the four different assemblies available for the A. deanei genome (GenBank ID 14191) ( Table S1 ).…”

Section: Are the Trypanososmatids´ Calpains Proteolytically Activementioning

confidence: 99%

“…Since these enzymes are involved in crucial physiological roles in mammals, their deregulated activity is implicated in several pathophysiological processes, especially in fibrotic diseases and neurological disorders [ 7 ]. Moreover, trypanosomatids harbor a large and diverse family of calpain sequences in their genomes, comprising a wide range of associated domains, differential gene expression among life-cycle forms, and ubiquitous distribution in the parasite cell body [ 8 , 9 , 10 , 11 ]. However, due to the difficulties in assaying calpain activity in these microorganisms, there is still an open question: are trypanosomatid calpains proteolytically active?…”

Section: Introductionmentioning

confidence: 99%

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The Diverse Calpain Family in Trypanosomatidae: Functional Proteins Devoid of Proteolytic Activity?

Ennes-Vidal

Branquinha

Santos

et al. 2021

Cells

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Calpains are calcium-dependent cysteine peptidases that were originally described in mammals and, thereafter, their homologues were identified in almost all known living organisms. The deregulated activity of these peptidases is associated with several pathologies and, consequently, huge efforts have been made to identify selective inhibitors. Trypanosomatids, responsible for life-threatening human diseases, possess a large and diverse family of calpain sequences in their genomes. Considering that the current therapy to treat trypanosomatid diseases is limited to a handful of drugs that suffer from unacceptable toxicity, tough administration routes, like parenteral, and increasing treatment failures, a repurposed approach with calpain inhibitors could be a shortcut to successful chemotherapy. However, there is a general lack of knowledge about calpain functions in these parasites and, currently, the proteolytic activity of these proteins is still an open question. Here, we highlight the current research and perspectives on trypanosomatid calpains, overview calpain description in these organisms, and explore the potential of targeting the calpain system as a therapeutic strategy. This review gathers the current knowledge about this fascinating family of peptidases as well as insights into the puzzle: are we unable to measure calpain activity in trypanosomatids, or are the functions of these proteins devoid of proteolytic activity in these parasites?

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Section: Calpain Superfamily Expansion In Trypanosomatid Genomes Amentioning

confidence: 99%

Section: Calpain Superfamily Expansion In Trypanosomatid Genomes Amentioning

confidence: 99%

Section: Calpain Superfamily Expansion In Trypanosomatid Genomes Amentioning

confidence: 99%

Section: Are the Trypanososmatids´ Calpains Proteolytically Activementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The Diverse Calpain Family in Trypanosomatidae: Functional Proteins Devoid of Proteolytic Activity?

Ennes-Vidal

Branquinha

Santos

et al. 2021

Cells

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Proteolytic inhibitors as alternative medicines to treat trypanosomatid-caused diseases: experience with calpain inhibitors

Ennes-Vidal

Santos

Branquinha

et al. 2022

Mem. Inst. Oswaldo Cruz

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The treatment for tropical neglected diseases, such as Chagas disease (CD) and leishmaniasis, is extremely limited to a handful of drugs that suffer from unacceptable toxicity, tough administration routes, like parenteral, and increasing treatment failures due to the parasite resistance. Consequently, there is urgency for the development of new therapeutic options to treat such diseases. Since peptidases from these parasites are responsible for crucial functions in their biology, these molecules have been explored as alternative targets. In this context, a myriad of proteolytic inhibitors has been developed against calciumdependent cysteine-type peptidases, collectively called calpains, which are implicated in several human pathophysiological diseases. These molecules are highly expanded in the genome of trypanosomatids and they have been reported participating in several parasite biological processes. In the present perspective, we discuss our almost two decades of experience employing the calpain inhibitors as an interesting shortcut to a possible repurpose strategy to treat CD and leishmaniasis.

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Oral infectivity through carnivorism in murine model of Trypanosoma cruzi infection

Torres,

Contreras,

Gutiérrez

et al. 2024

Front. Cell. Infect. Microbiol.

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IntroductionOral transmission of T. cruzi is probably the most frequent transmission mechanism in wild animals. This observation led to the hypothesis that consuming raw or undercooked meat from animals infected with T. cruzi may be responsible for transmitting the infection. Therefore, the general objective of this study was to investigate host-pathogen interactions between the parasite and gastric mucosa and the role of meat consumption from infected animals in the oral transmission of T. cruzi.MethodsCell infectivity assays were performed on AGS cells in the presence or absence of mucin, and the roles of pepsin and acidic pH were determined. Moreover, groups of five female Balb/c mice were fed with muscle tissue obtained from mice in the acute phase of infection by the clone H510 C8C3hvir of T. cruzi, and the infection of the fed mice was monitored by a parasitemia curve. Similarly, we assessed the infective capacity of T. cruzi trypomastigotes and amastigotes by infecting groups of five mice Balb/c females, which were infected orally using a nasogastric probe, and the infection was monitored by a parasitemia curve. Finally, different trypomastigote and amastigote inoculums were used to determine their infective capacities. Adhesion assays of T. cruzi proteins to AGS stomach cells were performed, and the adhered proteins were detected by western blotting using monoclonal or polyclonal antibodies and by LC-MS/MS and bioinformatics analysis. ResultsTrypomastigote migration in the presence of mucin was reduced by approximately 30%, whereas in the presence of mucin and pepsin at pH 3.5, only a small proportion of parasites were able to migrate (∼6%). Similarly, the ability of TCTs to infect AGS cells in the presence of mucin is reduced by approximately 20%. In all cases, 60–100% of the animals were fed meat from mice infected in the acute phase or infected with trypomastigotes or amastigotes developed high parasitemia, and 80% died around day 40 post-infection. The adhesion assay showed that cruzipain is a molecule of trypomastigotes and amastigotes that binds to AGS cells. LC-MS/MS and bioinformatics analysis, also confirmed that transialidase, cysteine proteinases, and gp63 may be involved in TCTs attachment or invasion of human stomach cells because they can potentially interact with different proteins in the human stomach mucosa. In addition, several human gastric mucins have cysteine protease cleavage sites. DiscussionThen, under our experimental conditions, consuming meat from infected animals in the acute phase allows the T. cruzi infection. Similarly, trypomastigotes and amastigotes could infect mice when administered orally, whereas cysteinyl proteinases and trans-sialidase appear to be relevant molecules in this infective process.

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Expression and cellular localisation of Trypanosoma cruzi calpains

Cited by 3 publications

References 31 publications

The Diverse Calpain Family in Trypanosomatidae: Functional Proteins Devoid of Proteolytic Activity?

The Diverse Calpain Family in Trypanosomatidae: Functional Proteins Devoid of Proteolytic Activity?

Proteolytic inhibitors as alternative medicines to treat trypanosomatid-caused diseases: experience with calpain inhibitors

Oral infectivity through carnivorism in murine model of Trypanosoma cruzi infection

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