Expression and One-Step Purification of recombinant Arabidopsis thaliana Frataxin homolog (AtFH)..

MALIANDI, M. V.; BUSI, M. V.; CLEMENTE, M.; EDUARDO JULIAN ZABALETA; ARAYA, A; GÓMEZ-CASATI, D.

PROTEIN EXPRESSION AND PURIFICATION

Springer

Año: 2007 vol. 51 p. 157 - 161

1046-5928

Frataxin, a nuclear-encoded mitochondrial protein, has been proposed to participate in Fe-S cluster assembly, mitochondrial energy metabolism, respiration and iron homeostasis. However, its precise function remains elusive. Frataxin is highly conserved in living organisms with no major structural changes, in particular at the C-terminal protein domain, suggesting that it plays a key function in all organisms. Recently, a plant gene, AtFH, with significant homology to other members of the frataxin family has been described. To gain insight on the frataxin role in plants, the frataxin domain was expressed in Escherichia coli BL21-CodonPlus (DE3)-RIL cells and chromatographed on Ni-chelating column. A single protein band of about 15 kDa, as assessed by SDS-PAGE, was obtained with 99% purity by one-step procedure. Furthermore, the addition of the purified protein to a mixture containing Fe(II) and H2O2 attenuate the Fenton reaction. The simply procedure described here may be useful to obtain large quantities of AtFH protein for further studies of its structure and function.