Functional characterization and subcellular distribution of two recombinant cytosolic HSP70 isoforms from Entamoeba histolytica under normal and stress conditions
Por:
Santos, Fabiola, Marcial-Quino, Jaime, Gomez-Manzo, Saul, Enriquez-Flores, Sergio, Nequiz-Avendano, Mario, Cortes, Azucena, De la Luz Leon-Avila, Gloria, Saavedra, Emma, Perez-Tamayo, Ruy, Olivos-Garcia, Alfonso
Publicada:
1 abr 2020
Resumen:
Amoebiasis is a human intestinal disease caused by the parasite
Entamoeba histolytica. It has been previously demonstrated that E.
histolytica heat shock protein 70 (EhHSP70) plays an important role in
amoebic pathogenicity by protecting the parasite from the dangerous
effects of oxidative and nitrosative stresses. Despite its relevance,
this protein has not yet been characterized. In this study, the EhHSP70
genes were cloned, and the two recombinant EhHSP70 proteins were
expressed, purifying and biochemically characterized. Additionally,
after being subjected to some host stressors, the intracellular
distribution of the proteins in the parasite was documented. Two amoebic
HSP70 isoforms, EhHSP70-A and EhHSP70-B, with 637 and 656 amino acids,
respectively, were identified. Kinetic parameters of ATP hydrolysis
showed low rates, which were in accordance with those of the HSP70
family members. Circular dichroism analysis showed differences in their
secondary structures but similarities in their thermal stability.
Immunocytochemistry in trophozoites detected EhHSP70 in the nuclei and
cytoplasm as well as a slight overexpression when the parasites were
subjected to oxidants and heat. The structural differences of amoebic
HSP70s with their human counterparts may be used to design specific
inhibitors to treat human amoebiasis.
Filiaciones:
Santos, Fabiola:
Univ Nacl Autonoma Mexico, Fac Med, Unidad Invest Med Expt, Mexico City 04510, DF, Mexico
Inst Politecn Nacl, Escuela Nacl Ciencias Biol, Mexico City 11340, DF, Mexico
Marcial-Quino, Jaime:
Inst Nacl Pediat, Lab Bioquim Genet, Secretaria Salud, Mexico City 04530, DF, Mexico
Gomez-Manzo, Saul:
Inst Nacl Pediat, Lab Bioquim Genet, Secretaria Salud, Mexico City 04530, DF, Mexico
Enriquez-Flores, Sergio:
Inst Nacl Pediat, Grp Invest Biomol & Salud Infantil, Secretaria Salud, Lab Errores Innatos Metab & Tamiz, Mexico City 04530, DF, Mexico
Nequiz-Avendano, Mario:
Univ Nacl Autonoma Mexico, Fac Med, Unidad Invest Med Expt, Mexico City 04510, DF, Mexico
Cortes, Azucena:
Univ Nacl Autonoma Mexico, Fac Med, Unidad Invest Med Expt, Mexico City 04510, DF, Mexico
De la Luz Leon-Avila, Gloria:
Inst Politecn Nacl, Escuela Nacl Ciencias Biol, Dept Zool, Mexico City 11340, DF, Mexico
Saavedra, Emma:
Inst Nacl Cardiol Ignacio Chavez, Dept Bioquim, Secretaria Salud, Mexico City 14080, DF, Mexico
Perez-Tamayo, Ruy:
Univ Nacl Autonoma Mexico, Fac Med, Unidad Invest Med Expt, Mexico City 04510, DF, Mexico
Olivos-Garcia, Alfonso:
Univ Nacl Autonoma Mexico, Fac Med, Unidad Invest Med Expt, Mexico City 04510, DF, Mexico
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