A monoclonal antibody against a Leishmania mexicana COX-like enzymatic activity also recognizes similar proteins in different protozoa of clinical importance

Hernández‐Ramírez, Verónica Ivonne; Estrada-Figueroa, Luis Alberto; Medina, Yolanda; Lizarazo-Taborda, Mélida R; Toledo-Leyva, Alfredo; Osorio-Trujillo, Juan Carlos; Morales-Mora, Daniel; Talamás-Rohana, Patricia

doi:10.1007/s00436-022-07746-7

Cited by 2 publications

(5 citation statements)

References 42 publications

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“…Furthermore, in recent years, COX activity has been determined in different parasites, proposing that part of the PGs in the infection microenvironment could be synthesized by the parasites. Hernández-Ramírez et al ( 2023), demonstrated COX activity in Naegleria fowleri, Entamoeba histolytica, Giardia duodenalis, Acanthamoeba castellanii, and Trypanosoma cruzi [17]. The presence of prostaglandins in Acanthamoeba spp.…”

Section: Discussionmentioning

confidence: 98%

“…Furthermore, Hernández-Ramírez et al (2023) reported a D12 mAb against the Gp63 protein of Leishmania mexicana monoclonal antibody, immunodetecting a COX-like protein in A. castellanii trophozoites [17]. In E. histolytica, the leishmanolysin homologue gene (E. histolytica MSP-1) has been reported, which encodes a surface metalloprotease involved in the regulation of amoebic adhesion, with additional effects on cell motility, the destruction of cell monolayers, and phagocytosis [60].…”

Section: Discussionmentioning

confidence: 99%

“…It is important to highlight that the areas of greatest intensity were observed in the pseudopodia of the trophozoites, areas of adhesion of these amoebae. The recognition of leishmanolysin-like proteins evidenced by surface labeling indicates similarities with other parasites [17]. It remains to be determined whether COX activity is also part of the functions of the same molecule, as well as enzymatic activity, specifically metalloprotease, and adhesin.…”

Section: Discussionmentioning

confidence: 99%

“…To determine whether the amoeba under study produced leishmanolysin-like proteins, that is, proteins with adhesin [16], protease, and cyclooxygenase (COX) activities [17], trophozoites and EVs were processed for confocal microscopy, as previously described, using mouse polyclonal antibody anti-GP63 leishmanolysin-like proteins (1:600) (USBiologid Life Science, Swampscott, MA, USA) as the primary antibody and anti-mouse IgG mAb (H+L) FITC antibody (1:600) (Invitrogen TM) as secondary antibody. Samples were observed using a Carl Zeiss LSM 700 confocal microscope.…”

Section: Leishmanolysin-like Protein Detection In a Culbertsoni Troph...mentioning

confidence: 99%

“…In addition, a putative Leishmanolysin gene of Acanthamoeba castellanii str. Neff (XP_004337275.1), a homologous protein to Leishmanolysin, GP63 from Leishmania, with adhesin [16], protease, and cyclooxygenase (COX) activities [17], has been described. Once firmly adhered, the amoebae migrate towards the cell junctions, and by mechanical and/or enzymatic effects trophozoites, penetrate gradually, introducing their acanthopods between the intercellular junctions [8,18,19], until invading and altering deeper layers and the target tissue architecture [9,10], without inducing the lysis or cytopathic effect of surrounding cells, or even without destabilizing its cytoskeleton [8].…”

Section: Introductionmentioning

confidence: 99%

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Extracellular Vesicles Secreted by Acanthamoeba culbertsoni Have COX and Proteolytic Activity and Induce Hemolysis

Sierra-López,

Castelan-Ramírez,

Hernández-Martínez

et al. 2023

Microorganisms

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Several species of Acanthamoeba genus are potential pathogens and etiological agents of several diseases. The pathogenic mechanisms carried out by these amoebae in different target tissues have been documented, evidencing the relevant role of contact-dependent mechanisms. With the purpose of describing the pathogenic processes carried out by these protozoans more precisely, we considered it important to determine the emission of extracellular vesicles (EVs) as part of the contact-independent pathogenicity mechanisms of A. culbertsoni, a highly pathogenic strain. Through transmission electronic microscopy (TEM) and nanoparticle tracking analysis (NTA), EVs were characterized. EVs showed lipid membrane and a size between 60 and 855 nm. The secretion of large vesicles was corroborated by confocal and TEM microscopy. The SDS-PAGE of EVs showed proteins of 45 to 200 kDa. Antigenic recognition was determined by Western Blot, and the internalization of EVs by trophozoites was observed through Dil-labeled EVs. In addition, some EVs biological characteristics were determined, such as proteolytic, hemolytic and COX activity. Furthermore, we highlighted the presence of leishmanolysin in trophozites and EVs. These results suggest that EVs are part of a contact-independent mechanism, which, together with contact-dependent ones, allow for a better understanding of the pathogenicity carried out by Acanthamoeba culbertsoni.

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Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Leishmanolysin-like Protein Detection In a Culbertsoni Troph...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Extracellular Vesicles Secreted by Acanthamoeba culbertsoni Have COX and Proteolytic Activity and Induce Hemolysis

Sierra-López,

Castelan-Ramírez,

Hernández-Martínez

et al. 2023

Microorganisms

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Exploration of the Binding Site of Arachidonic Acid in gp63 of Leishmania mexicana and in Orthologous Proteins in Clinically Important Parasites

Hernández-Ramírez,

Matus-Meza,

Oviedo

et al. 2024

Pathogens

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Recently, we published that the monoclonal antibody (D12 mAb) recognizes gp63 of L. mexicana, and it is responsible for COX activity. This D12 mAb exhibited cross-reactivity with Trypanosoma cruzi, Entamoeba histolytica, Acanthamoeba castellanii, and Naegleria fowleri. COX activity assays performed in these parasites suggested the potential presence of such enzymatic activity. In our investigation, we confirmed that wild-type recombinant gp63 exhibits COX-like activity, in contrast to a mutated recombinant gp63 variant. Consequently, our objective was to identify sequences orthologous to gp63 and subsequently analyze the binding of arachidonic acid (AA) to the putative active sites of these proteins. Given the absence of a crystallized structure for this protein in the Protein Data Bank (PDB), it was imperative to first obtain a three-dimensional structure by homology modeling, using leishmanolysin from Leishmania major (PDB ID: LML1) as a template in the Swiss model database. The results obtained through molecular docking simulations revealed the primary interactions of AA close to the Zinc atom present in the catalytic site of gp63-like molecules of several parasites, predominantly mediated by hydrogen bonds with HIS264, HIS268 and HIS334. Furthermore, COX activity was evaluated in commensal species such as E. dispar and during the encystment process of E. invadens.

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A monoclonal antibody against a Leishmania mexicana COX-like enzymatic activity also recognizes similar proteins in different protozoa of clinical importance

Cited by 2 publications

References 42 publications

Extracellular Vesicles Secreted by Acanthamoeba culbertsoni Have COX and Proteolytic Activity and Induce Hemolysis

Extracellular Vesicles Secreted by Acanthamoeba culbertsoni Have COX and Proteolytic Activity and Induce Hemolysis

Exploration of the Binding Site of Arachidonic Acid in gp63 of Leishmania mexicana and in Orthologous Proteins in Clinically Important Parasites

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