Biochemical and Molecular Characterization of a Novel Cu/Zn Superoxide Dismutase from Amaranthus hypochondriacus L.: an Intrinsically Disordered Protein
Por:
Montero-Morán G.M., Sampedro J.G., Saab-Rincón G., Cervantes-González M.A., Huerta-Ocampo J.Á., De León-Rodríguez A., de la Rosa, AP
Publicada:
1 ago 2015
Resumen:
A novel Cu/ZnSOD from Amaranthus hypochondriacus was cloned, expressed, and characterized. Nucleotide sequence analysis showed an open reading frame (ORF) of 456 bp, which was predicted to encode a 15.6-kDa molecular weight protein with a pI of 5.4. Structural analysis showed highly conserved amino acid residues involved in Cu/Zn binding. Recombinant amaranth superoxide dismutase (rAhSOD) displayed more than 50 % of catalytic activity after incubation at 100 °C for 30 min. In silico analysis of Amaranthus hypochondriacus SOD (AhSOD) amino acid sequence for globularity and disorder suggested that this protein is mainly disordered; this was confirmed by circular dichroism, which showed the lack of secondary structure. Intrinsic fluorescence studies showed that rAhSOD undergoes conformational changes in two steps by the presence of Cu/Zn, which indicates the presence of two binding sites displaying different affinities for metals ions. Our results show that AhSOD could be classified as an intrinsically disordered protein (IDP) that is folded when metals are bound and with high thermal stability. © 2015, Springer Science+Business Media New York.
Filiaciones:
Montero-Morán G.M.:
División Biología Molecular, IPICyT, Instituto Potosino de Investigación Científica y Tecnológica, Camino a la Presa San José 2055, Col. Lomas 4a. Sección, San Luis Potosí, SLP 78216, Mexico
Sampedro J.G.:
Instituto de Física, Universidad Autónoma de San Luis Potosí, Manuel Nava 6, zona Universitaria, San Luis Potosí, SLP 78290, Mexico
Saab-Rincón G.:
UNAM, Inst Biotechnol, Dept Ingn Celular & Biocatalisis, Cuernavaca 62271, Morelos, Mexico
Cervantes-González M.A.:
División Biología Molecular, IPICyT, Instituto Potosino de Investigación Científica y Tecnológica, Camino a la Presa San José 2055, Col. Lomas 4a. Sección, San Luis Potosí, SLP 78216, Mexico
Huerta-Ocampo J.Á.:
División Biología Molecular, IPICyT, Instituto Potosino de Investigación Científica y Tecnológica, Camino a la Presa San José 2055, Col. Lomas 4a. Sección, San Luis Potosí, SLP 78216, Mexico
De León-Rodríguez A.:
División Biología Molecular, IPICyT, Instituto Potosino de Investigación Científica y Tecnológica, Camino a la Presa San José 2055, Col. Lomas 4a. Sección, San Luis Potosí, SLP 78216, Mexico
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