Fig. 4
Different preparations of PorACur visualized on a 16.5% SDS-PAGE containing tricine. The SDS-PAGE was stained with silver. Lanes 1 and 4, Low molecular mass protein ladder. Lane 2, Purified PorACur with C-terminal His8-tag expressed in C. glutamicum ATCC 13032 ∆HA, Lane 3, Purified PorACur from preparative gel without tag expressed in C. glutamicum ATCC 13032 ∆HA, Lane 5, Purified CUR_1715 without GST-tag (around 9 KDa) expressed in E. coli BL21 DE3 (Omp8). The purified proteins with and without tags were reconstituted into lipid bilayer membranes. Only protein of lane 3 shows channel-formed activity in lipid bilayers. The channel-forming protein, named PorACur (coded by cur_1714 gene), was identified in the known genome of C. urealyticum by its similar chromosomal localization to known porH and porA genes of other Corynebacterium strains (see Fig. 3). The arrows indicate the location of the corresponding proteins