Mycobacterial CoaE domain organization and protein expression.
A, The mycobacterial CoaE has a 196 amino acid (a.a.) N-terminal dephosphocoenzyme A kinase and a 211 a.a. C-terminal domain of unknown function, the UPF0157. B, elution profile of hexahistidine tagged mycobacterial CoaE purified by affinity chromatography as collected fractions loaded on a 12% SDS-PAGE, M, protein molecular weight marker; P, cell pellet obtained post sonication and centrifugation; L, supernatant obtained post centrifugation of the sonicate, loaded onto the column; FT, Flow through from the column upon loading the supernatant; Wash, Fractions collected during column washing with resuspension buffer; Elution, the elution profile represented by fractions collected during elution with buffer containing 100 mM imidazole. C, confirmation of the clone by Western blotting using anti-Histidine antibodies against the recombinant His-tagged CoaE (Lane 3) and His-tagged Plasmodium falciparum FabI as a positive control (Lane2). Lane 1 carries the pre-stained molecular weight marker.