Transcriptome-Based Identification of a Functional Fasciola hepatica Carboxylesterase B
Por:
Pedroza-Gomez, Yaretzi J., Cossio-Bayugar, Raquel, Aguilar-Diaz, Hugo, Scarcella, Silvana, Reynaud, Enrique, Sanchez-Carbente, Maria del Rayo, Narvaez-Padilla, Veronica, Miranda-Miranda, Estefan
Publicada:
1 nov 2021
Resumen:
Bioinformatics analysis of the complete transcriptome of Fasciola
hepatica, identified a total of ten putative carboxylesterase
transcripts, including a 3146 bp mRNA transcript coding a 2205 bp open
reading frame that translates into a protein of 735 amino acids,
resulting in a predicted protein mass of 83.5 kDa and a putative
carboxylesterase B enzyme. The gene coding for this enzyme was found in
two reported F. hepatica complete genomes stretching 23,230 bp,
containing two exons of 1282 and 1864 bp, respectively, as well as a
20,084 bp intron between the exons. The enzymatic activity was
experimentally assayed on F. hepatica protein extracts by SDS-PAGE
zymograms using synthetic chromogenic substrates, confirming both the
theoretical molecular weight and carboxylesterase enzymatic activity.
Further bioinformatics predicted that this enzyme is an integral
component of the cellular membrane that should be active as a 167 kDa
homodimer complex and polyacrylamide gel electrophoresis (PAGE)
zymograms experiments confirmed the analysis. Additional bioinformatics
analysis showed that DNA sequences that code for this particular enzyme
are highly conserved in other parasitic trematodes, although they are
labeled hypothetical proteins.
Filiaciones:
Pedroza-Gomez, Yaretzi J.:
Centro Nacional de Investigaciones Disciplinarias en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales Agrícolas y Pecuarias (INIFAP), Boulevard Cuauhnahuac No. 8534, Jiutepec, Morelos, 62550, Mexico
Cossio-Bayugar, Raquel:
Centro Nacional de Investigaciones Disciplinarias en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales Agrícolas y Pecuarias (INIFAP), Boulevard Cuauhnahuac No. 8534, Jiutepec, Morelos, 62550, Mexico
Aguilar-Diaz, Hugo:
Centro Nacional de Investigaciones Disciplinarias en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales Agrícolas y Pecuarias (INIFAP), Boulevard Cuauhnahuac No. 8534, Jiutepec, Morelos, 62550, Mexico
Scarcella, Silvana:
Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires, Campus Universitario, Tandil, 7000, Argentina
Reynaud, Enrique:
Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Avenida Universidad, 2001, Apartado Postal 510–3, Cuernavaca, Morelos, 62210, Mexico
Sanchez-Carbente, Maria del Rayo:
Centro de Investigación en Biotecnología de la Universidad Autónoma del Estado de Morelos, Av. Universidad 1001, Cuernavaca, Morelos, 62209, Mexico
Narvaez-Padilla, Veronica:
Centro de Investigación en Dinámica Celular de la Universidad Autónoma del Estado de Morelos, Cuernavaca Morelos México, Av. Universidad 1001, Chamilpa, Cuernavaca, Morelos, 62210, Mexico
Miranda-Miranda, Estefan:
Centro Nacional de Investigaciones Disciplinarias en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales Agrícolas y Pecuarias (INIFAP), Boulevard Cuauhnahuac No. 8534, Jiutepec, Morelos, 62550, Mexico
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