Expression, Purification and Crystallization of Pore Mutants of Ammonium Transport Protein 1 From Archaeoglobus Fulgidus

06/05/2020

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by   Daniel Cebo, et al.

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Ammonium transport proteins are highly conserved families of integral membrane proteins present in all domains of life. Functional studies with members of this family have yielded controversial results with respect to the chemical identity of the transported species (NH4+ or NH3) as well as to the classification of the proteins as channels or secondary active transporters. In this work different point mutations of Ammonium transport protein 1 gene (Amt-1) have been produced by site-directed mutagenesis. Then heterologous overexpression in Escherichia coli has been used to produce proteins and they have been purified to electrophoretic homogeneity. Crystals of Amt-1 have been obtained by sitting-drop vapor diffusion method.