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. 2015 May 19;5:9747. doi: 10.1038/srep09747

Figure 4. Analysis of partially purified WT Eco94GmrSD and mutant proteins D507A, H508A, C517A, N522A, N528A, N535A, E271A/E273A, E278A/K280A on SDS-PAGE and endonuclease activity assays for the mutant enzymes on T4 DNA.

Figure 4

(A). SDS-PAGE analysis of WT and mutant proteins. Left panel: WT and mutant proteins (D507A, H508A, N522A, and N535A) purified by nickel-NTA agarose and heparin HP columns (purified D507A showed aberrant migration). Right panel: WT, E271A/E273A, E278A/K280A, C517A and N528A proteins. (B and C). Endonuclease activity assay for WT and GmrSD variants D507A, H508A, C517A, N522A, N528A, N535A on T4 DNA. The amount of input protein was 0.5 μg, 1 μg, and 2 μg, respectively in digestion of 1 μg T4 DNA. For the double mutants E271A/E273A and E278A/K280A, the amount of input protein was 0.5 μg, 1 μg, 1.5 μg and 2 μg, respectively in digestion of 1 μg T4 DNA.