Metallopeptidases of<i>Toxoplasma gondii</i>:<i>in silico</i>identification and gene expression

Escotte-Binet, Sandie; Huguenin, A; Aubert, Dominique; Martin, Anne-Pascaline; Kaltenbach, Matthieu L.; Florent, Isabelle; Villena, Isabelle

doi:10.1051/parasite/2018025

Cited by 17 publications

(15 citation statements)

References 69 publications

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“…The domain I contains an active site characterized by the zinc-binding motif HYLEH, while domains II and IV have homology to the M16A inactive catalytic domains of insulin-like proteases, suggests that INS-15 is a functional M16A protease [24]. Pf SPP, an M16C protein in P. falciparum , cleaves the transit peptide of plastid-targeted proteins [21,25,26]. Falcilysin is another M16C protein in P. falciparum and was shown to degrade transit peptide in the apicoplast [20].…”

Section: Discussionmentioning

confidence: 99%

Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum

Guo

et al. 2019

Microorganisms

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Cryptosporidium parvum is a protozoan parasite that can cause moderate-to-severe diarrhea. Insulinase-like proteases (INS) are one of the largest protein families within the small proteome of the pathogen. However, their roles in C. parvum biology remain un-elucidated. In this study, a member of the protein family, INS-15 of C. parvum encoded by cgd3_4260, was cloned, expressed and characterized to understand its function. INS-15 and its domain I were expressed in Escherichia coli and polyclonal antibodies against the domain I and one specific polypeptide were prepared in rabbits. The role of INS-15 protein in the C. parvum invasion was preliminarily studied. Recombinant INS-15 protein and its domain I were successfully expressed in E. coli, together with various degraded products. The cgd3_4260 gene had a peak expression at 2 h of in vitro C. parvum culture, while the INS-15 protein was expressed in the mid-anterior region of sporozoites and the area of merozoites opposite to the nucleus. Anti-INS-15 domain I antibodies reduced the invasion of C. parvum sporozoites by over 40%. The anterior location of INS-15 in invasion stages and partial reduction of in vitro growth indicate that INS-15 plays some roles in the invasion or early development of C. parvum.

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Section: Discussionmentioning

confidence: 99%

Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum

Guo

et al. 2019

Microorganisms

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“…Metalloproteases are widespread in biology and they have been classified into 16 clans that are summarized in the MEROPS database (https://www.ebi.ac.uk/merops/). M16 metalloproteases are characterized by the presence of a zinc binding motif consisting of the sequence HXXEH (22). M16A and M16C family member contain four domains, only one of which contains the active catalytic site (22).…”

Section: Discussionmentioning

confidence: 99%

“…M16 metalloproteases are characterized by the presence of a zinc binding motif consisting of the sequence HXXEH (22). M16A and M16C family member contain four domains, only one of which contains the active catalytic site (22). The best known M16 protease is human insulinase that cleaves variety type of small peptides in different type of cells, consistent with multiple roles of this enzyme (31).…”

Section: Discussionmentioning

confidence: 99%

“…M16 metalloproteases typically bind zinc as part of their active site (HXXEH) and they cleave short polypeptides, the size of which is constrained by a conserved small barrel fold that forms the catalytic chamber (21). The apicomplexan parasite Toxoplasma gondii contains ~ 50 metalloproteases, including 11 members of the M16 clan (22), several of which are found in secretory organelles implicated in host cell interactions (23, 24). In Plasmodium falciparum the M16 metalloprotease falcilysin participates in both hemoglobin degradation and in processing of transit peptides for apicoplast proteins (25, 26).…”

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confidence: 99%

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Insulinase like protease 1 contributes to macrogamont formation inCryptosporidium parvum

Xu¹,

Feng²,

Xiao³

et al. 2020

Preprint

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The apicomplexan parasite Cryptosporidium parvum contains an expanded family of 22 insulinase like proteases (INS), a feature that contrasts with their otherwise streamlined genome. Here we examined the function of INS1, which is most similar to the human insulinase protease that cleaves a variety of small peptide substrates. INS1 is a M16A clan member and contains a signal peptide, an N-terminal domain with the HxxEH active site, followed by three inactive domains. Unlike previously studied C. parvum INS proteins that are expressed in sporozoites and during merogony, INS1 was expressed exclusively in macrogamonts, where it was localized in small cytoplasmic vesicles. Although INS1 did not colocalize with the oocyst wall protein recognized by the antibody OW50, immune-electron microscopy indicated that INS1 resides in small vesicles in the secretory system. Notably, these small INS1 positive vesicles often subtend large vacuoles resembling wall forming bodies, which contain precursors for oocyst wall formation. Genetic deletion of INS1, or replacement with an active site mutant, resulted in lower formation of macrogamonts in vitro and reduced oocyst shedding in vivo. Our findings reveal that INS1 functions in formation or maturation of macrogamonts and that its loss results in attenuated virulence in immunocompromised mice.ImportanceCryptosporidiosis is a debilitating diarrheal disease in young children in developing countries. Absence of effective treatments or vaccines makes this infection very difficult to manage in susceptible populations. Although the oral dose of oocysts needed to cause infection is low, infected individuals shed very high numbers of oocysts, hence readily contaminating the environment. Our studies demonstrate that the protease INS1 is important for formation of female sexual stages and that in its absence, parasites produce fewer oocysts and are attenuated in immunocompromised mice. These findings suggest that mutants lacking INS1, or related proteases, may be useful for producing attenuated vaccines to induce immunity without causing disease.

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“…[3][4][5] The driving forces that rule the interaction between two proteins lie in certain regions hidden in the primary structure of the proteins; therefore, the discovery of these regions in pathogenic proteins is essential to implement future therapies that could block pathogen infections. 6 Most computational tools to infer interacting regions in proteins require three-dimensional (3D) structure information from protein complexes; unfavorably, the majority of PPI complexes have no crystallography information, which is why inferences for interaction regions should be predicted from the primary structure of proteins stored in the pathogen databases. 7 Many of the interaction regions in proteins rely on their short linear motifs (SLiMs).…”

Section: Introductionmentioning

confidence: 99%

Time-Frequency Approach Applied to Finding Interaction Regions in Pathogenic Proteins

Arenas

Arango-Plaza

Arenas

et al. 2019

Bioinform Biol Insights

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Protein-protein interactions govern all molecular processes for living organisms, even those involved in pathogen infection. Pathogens such as virus, bacteria, and parasites contain proteins that help the pathogen to attach, penetrate, and settle inside the target cell. Thus, it is necessary to know the regions in pathogenic proteins that interact with host cell receptors. Currently, powerful pathogen databases are available and many pathogenic proteins have been recognized, but many pathogenic proteins have not been characterized. This work developed a program in MATLAB environment based on the time-frequency analysis to recognize important sites in proteins. Our program highlights the highest energy patches in proteins from their time-frequency distribution and matches the corresponding frequency. We sought to know if this approach is able to recognize stretches residues related to interaction. Our approach was applied to five study cases from pathogenic co-crystallized structures that have been well characterized. We searched the frequencies that characterize interaction regions in pathogenic proteins and with this information tried to identify new interaction patches in either paralogs or orthologs. We found that our program generates a well-interpretable graphic under several descriptors that can show important regions in proteins even those related to interaction. We propose that this MATLAB program could be used as a tool to explore outstanding regions in uncharacterized proteins.

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Metallopeptidases ofToxoplasma gondii:in silicoidentification and gene expression

Cited by 17 publications

References 69 publications

Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum

Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum

Insulinase like protease 1 contributes to macrogamont formation inCryptosporidium parvum

Time-Frequency Approach Applied to Finding Interaction Regions in Pathogenic Proteins

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