Mostrar el registro sencillo del ítem
dc.contributor.author
Carmona Viglianco, Natalia Evelín
dc.contributor.author
Mascotti, María Laura
dc.contributor.author
Giannini, Fernando A.
dc.contributor.author
Baldoni, Hector Armando
dc.date.available
2024-01-12T09:52:10Z
dc.date.issued
2019
dc.identifier.citation
Improvement in the production of active cruzain for in vitro inhibitors screening assays; XXII Simposio Nacional de Química Orgánica; Mendoza; Argentina; 2019; so-75
dc.identifier.isbn
978-987-733-194-3
dc.identifier.uri
http://hdl.handle.net/11336/223462
dc.description.abstract
Chagas disease is a chronic systemic parasitic infection caused by the protozoan parasite Trypanosoma cruzi. A promising target for potential therapeutics against it is the major cysteine protease expressed by T. cruzi, cruzain. Cruzain is a member of the papain superfamily of cysteine proteases, and it is homologous to cathepsin-L. These enzymes degrade polypeptides and are characterized by having a common catalytic mechanism that involves a nucleophilic cysteine thiol in the catalytic triad. In the case of Chagas disease, cruzain is present in all life stages of the trypanosome and is implicated in cellular entry and digestion of immunoglobins. Inhibition studies of cruzain in cell models indicated prevention and reduction of infection.Previously, we demonstrated the expression of a recombinant cruzain in E. coliBL21(DE3) classic, pLys E, and Rosetta strains.1 However, most of the produced protein was present as protein aggregates. Optimization of culture conditions, including inductor concentrations and temperature shifts, allowed us to maximize the production of soluble active cruzain in concentration, yield and purity suitable for in vitro high throughput screening assays to identify potentially new and novel cruzain inhibitors by both kinetic and endpoint assays. Preliminary fluorometric assays showed that the mature cruzain cleaves the synthetic fluorogenic peptide substrate N-CBZ-Phe-Arg-7-amido-4-methylcoumarin under our experimental conditions. With the aim of identifying new chemical scaffolds leading to the development of novel drugs useful in the treatment ofChagas' disease, we offer this assay for joint research activities and collaborations.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Sociedad Argentina de Investigación
dc.rights
info:eu-repo/semantics/restrictedAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
CHAGAS
dc.subject
CRUZAIN
dc.subject
CRUZIPAIN
dc.subject
INHIBITORS
dc.subject.classification
Bioquímica y Biología Molecular
dc.subject.classification
Ciencias Biológicas
dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
dc.title
Improvement in the production of active cruzain for in vitro inhibitors screening assays
dc.type
info:eu-repo/semantics/publishedVersion
dc.type
info:eu-repo/semantics/conferenceObject
dc.type
info:ar-repo/semantics/documento de conferencia
dc.date.updated
2023-02-16T11:24:14Z
dc.journal.pagination
so-75
dc.journal.pais
Argentina
dc.journal.ciudad
Mendoza
dc.description.fil
Fil: Carmona Viglianco, Natalia Evelín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
dc.description.fil
Fil: Mascotti, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
dc.description.fil
Fil: Giannini, Fernando A.. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Área Química General e Inorgánica; Argentina
dc.description.fil
Fil: Baldoni, Hector Armando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Matemática Aplicada de San Luis "Prof. Ezio Marchi". Universidad Nacional de San Luis. Facultad de Ciencias Físico, Matemáticas y Naturales. Instituto de Matemática Aplicada de San Luis "Prof. Ezio Marchi"; Argentina
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.conicet.rol
Autor
dc.coverage
Nacional
dc.type.subtype
Simposio
dc.description.nombreEvento
XXII Simposio Nacional de Química Orgánica
dc.date.evento
2019-11-05
dc.description.ciudadEvento
Mendoza
dc.description.paisEvento
Argentina
dc.type.publicacion
Book
dc.description.institucionOrganizadora
Sociedad Argentina de Investigación en Química Orgánica
dc.source.libro
Libros de resúmenes: XXII Simposio Nacional de Química Orgánica
dc.date.eventoHasta
2019-11-08
dc.type
Simposio
Archivos asociados