Figure 2.

Disruption of Sumo1 sites K122/142 R did not alter cellular localization of Prdx6. (a) Schematic diagram of the evolutionary conserved Sumo1 motifs of Prdx6 spotted by SUMOplot and Prdx6 mutants fused to GFP plasmid as shown. SUMOplot, a web-based software program (http://sumosp.biocuckoo.org/online.php), was used to examine Sumo1 conjugation motif. Sequence alignment of human, mouse, rat, monkey and cow Prdx6 protein was conducted to identify evolutionary conserved Sumoylation motif (ClustalW). Lysine (K) residues are indicated in red bold letters. (b) Localization of WT and K122R or K142 R or K122/142 R mutant of Prdx6 fused to GFP plasmid. To examine if putative Sumoylation motifs K122 or K142 in Prdx6 protein predicted by Web-based analysis is indeed responsible for Sumo1 conjugation, K residue was changed to R by using SDM and tested for localization pattern for each plasmid. Immunofluorescence images showing localization of Prdx6 and its mutant forms K122R, K142 R and K122/142 R; cells transfected with pGFP-Prdx6WT (WT, upper left panel), mutant pGFP-Prdx6K122R (K122R, upper right panel), pGFP-Prdx6K142R (K142R, lower right panel), pGFP-Prdx6K122/142R (K122/142R, lower right panel) and fluorescence images of live cells were recorded after 24 h of transfection under inverted fluorescence microscope (Nikon Eclipse Ti-U)