Protein C-Terminal Labeling and Biotinylation Using Synthetic Peptide and Split-Intein
Figure 5
On-column production of ML protein.
A. The co-purification labeling process was monitored through SDS-PAGE analysis with Coomassie blue staining or fluorescence scanning as indicated. Lanes 1 and 2: total E. coli proteins before and after IPTG-induced expression of the precursor protein MINC, respectively. Lane 3: soluble fraction of the cell lysate of lane 2. Lane 4: proteins of lane 3 bound to chitin beads. Lane 5: proteins released from the chitin beads of lane 4 after an overnight incubation with the labeling peptide IC-L in the presence of TCEP. B. Amylose resin was incubated with (panels 1 and 2) or without (panels 3 and 4) the purified ML protein. Panels 1 and 3 are differential interference contrast images, while panels 2 and 4 are fluorescence images. C. SDS-PAGE analysis of the ML protein fractions eluted from the amylose resin of panels 3 and 4 of B using a maltose-containing elution buffer.