Characterization of proteolytic activities of Giardia lamblia with the ability to cleave His-tagged N-terminal sequences
Por:
de la Mora-de la Mora J.I., Enriquez-Flores, Sergio, Fernandez-Lainez, Cynthia, Gutierrez-Castrellon, Pedro, Olivos-Garcia, Alfonso, Gonzalez-Canto, Augusto, Hernandez, Roberto, Lujan, Hugo D., Garcia-Torres, Itzhel, Lopez-Velazquez, Gabriel
Publicada:
1 mar 2019
Resumen:
Giardia lamblia is one of the most common protozoan infectious agents in the world and is responsible for diarrheal disease and chronic postinfectious illness. During the host-parasite interaction, proteases are important molecules related to virulence, invasion, and colonization, not only for Giardia but also for other parasites. We aimed to characterize the cysteine protease activity detected in trophozoite lysates. This proteolytic activity showed the ability to cleave NH-terminal sequences with either a recognition sequence for a viral protease or a recognition sequence for thrombin. This cleavage activity was detected in nonencysting trophozoites and increased with the progression of encystation. This activity was also detected in excretion/secretion products of axenic trophozoites and in trophozoites cocultured with differentiated Caco-2 cells. Based on size exclusion chromatography, we obtained a fraction enriched in low- to medium-molecular-weight proteins that was capable of exerting this cleavage activity and aggregating human platelets. Finally, our results suggest that this proteolytic activity is shared with other protozoan parasites. © 2019 Elsevier B.V.
Filiaciones:
Grupo de Investigación en Biomoléculas y Salud Infantil, Laboratorio de EIMyT, Instituto Nacional de Pediatría, S.S., Mexico City, 04530, Mexico
Laboratorio de Errores Innatos del Metabolismo y Tamiz (EIMyT), Instituto Nacional de Pediatría, S.S., Mexico City, 04530, Mexico
Centro de Investigación Traslacional en salud Materno-Infantil. Hospital General Dr. Manuel Gea González, Mexico City, 14080, Mexico
Unidad de Investigación en Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de México, Mexico City, 04510, Mexico
Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Mexico City, 04510, Mexico
Laboratorio de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad Católica de Córdoba, Argentina
Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas (CONICET), Córdoba, Argentina
Inst Nacl Pediat, Lab EIMyT, Grp Invest Biomol & Salud Infantil, SS, Mexico City 04530, DF, Mexico
Inst Nacl Pediat, Lab Errores Innatos Metab & Tamiz EIMyT, SS, Mexico City 04530, DF, Mexico
Hosp Gen Dr Manuel Gea Gonzalez, Ctr Invest Traslac Salud Maternoinfantil, Mexico City 14080, DF, Mexico
Univ Nacl Autonoma Mexico, Fac Med, Unidad Invest Med Expt, Mexico City 04510, DF, Mexico
Univ Nacl Autonoma Mexico, Inst Invest Biomed, Dept Biol Mol & Biotecnol, Mexico City 04510, DF, Mexico
Univ Catolica Cordoba, Fac Med, Lab Bioquim & Biol Mol, Cordoba, Argentina
Consejo Nacl Invest Cient & Tecn, Ctr Invest & Desarrollo Inmunol & Enfermedades In, Cordoba, Argentina
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