Exploration of the Binding Site of Arachidonic Acid in gp63 of <i>Leishmania mexicana</i> and in Orthologous Proteins in Clinically Important Parasites
Verónica Ivonne Hernández-Ramírez,
Audifás-Salvador Matus-Meza,
Norma Oviedo,
Marco Antonio Magos-Castro,
Carlos Osorio-Trujillo,
Lizbeth Salazar-Villatoro,
Luis Alejandro Constantino-Jonapa,
Patricia Talamás-Rohana
Affiliations
Verónica Ivonne Hernández-Ramírez
Departamento de Infectómica y Patogénesis Molecular, CINVESTAV, Av. IPN No. 2508, Col. San Pedro Zacatenco, México City 07360, Mexico
Audifás-Salvador Matus-Meza
Department of Pharmaceutical Sciences, College of Pharmacy, University of Nebraska Medical Center, Omaha, NE 69198, USA
Norma Oviedo
Unidad de Investigación Médica en Inmunología e Infectología, Centro Médico Nacional La Raza, IMSS, Av. Jacarandas S/N, La Raza, Azcapotzalco, México City 02990, Mexico
Marco Antonio Magos-Castro
Departamento de Genética y Biología Molecular, CINVESTAV, Av. IPN No. 2508, Col. San Pedro Zacatenco, México City 07360, Mexico
Carlos Osorio-Trujillo
Departamento de Infectómica y Patogénesis Molecular, CINVESTAV, Av. IPN No. 2508, Col. San Pedro Zacatenco, México City 07360, Mexico
Lizbeth Salazar-Villatoro
Departamento de Infectómica y Patogénesis Molecular, CINVESTAV, Av. IPN No. 2508, Col. San Pedro Zacatenco, México City 07360, Mexico
Luis Alejandro Constantino-Jonapa
Unidad de Investigación UNAM-INC, División de Investigación, Facultad de Medicina, UNAM, Instituto Nacional de Cardiología Ignacio Chávez, México City 14080, Mexico
Patricia Talamás-Rohana
Departamento de Infectómica y Patogénesis Molecular, CINVESTAV, Av. IPN No. 2508, Col. San Pedro Zacatenco, México City 07360, Mexico
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.
