FIGURE 1.

In silico analyses and functional characterization of multifunctional protein PE_PGRS11. A, i, the presence of the phosphoglycerate mutase domain toward the C terminus of PE_PGRS11 protein upon the search against the domain database. Phosphoglycerate mutase matched with region 287–374 of PE_PGRS11 upon comparison with four Protein Data Bank structures, 1K6M, 1C81, 1TIP, and 1V7Q. All the active site residues of PE_PGRS11 (by active site prediction methods) were identical with these templates. ii, a three-dimensional model of the PE_PGRS11 phosphoglycerate mutase domain constructed on the basis of template 1K6M. iii, representation of the active site residues of phosphoglycerate mutase with 3-phosphoglyceric acid as a ligand upon automated docking simulations with the AutoDock 3.0 software suite using Silicon Graphics station Octane. B, a typical hyperbolic substrate saturation plot for the phosphoglycerate mutase enzyme activity of PE_PGRS11. The inset shows a Lineweaver-Burk plot to derive K_m and the V_max values for the phosphoglycerate mutase enzyme activity of PE_PGRS11. C, quantitative real time RT-PCR analysis of PE_PGRS11 or HspX transcripts assessed in the RNA isolated from M. tuberculosis grown under the indicated growth conditions. Error bars represent mean ± S.E. (n = 3). The data represent three independent experiments.