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. 2016 Oct 28;7:1677. doi: 10.3389/fmicb.2016.01677

Figure 2.

Figure 2

Size exclusion chromatography of PlmA indicates that it is a dimer. Pure recombinant PlmA (0.1 mg; details of production to be reported elsewhere; purity illustrated in the inset showing Coomassie-stained 12% SDS-PAGE gel; St, PageRuler prestained mass standards, from Thermo Scientific) was applied in 0.1 ml to a SuperdexTM 200 10/300 GL column (GE Healthcare) mounted on a ÄKTA FPLC system run at 0.3 ml/min of 50 mM Hepes, pH 7.5, 0.5 M NaCl, and 1 mM 2-mercaptoethanol, monitoring the optical absorption of the effluent at 280 nm (bottom graph). A semilogarithmic plot of mass relative to elution volume of protein standards is shown at the top. This plot was prepared with thyroglobulin, ferritin, β-amylase, bovine serum albumin, carbonic anhydrase, ribonuclease A, and cytochrome C, having respective masses (in kDa) of 669, 443, 200, 66.4, 29, 13.7, and 12.4. The open circle marks the elution position of the PlmA peak assuming that it is a dimer (sequence-deduced mass for the dimer, 72.7 kDa).