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![[Figure 4]](yt5033fig4mag.jpg)
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Figure 4
T-ag OBD triple mutants form ladders. Results of Cu(1,10-phenanthroline)2SO4 (CuP) catalyzed oxidation experiments electrophoresed on an SDS-polyacrylamide gel and stained with Coomassie Brilliant Blue. Lane 3 contains molecular-weight markers (250, 150, 100, 75, 50, 37, 25, 20, 15 and 10 kDa). Lanes 4–8 contain varying amounts of the triple mutant (3XM): 1x, 2x or 4x, where x = 1 mg ml−1. Lane 1 contains the reduced 3XM protein and lanes 2 and 4 show the 3XM protein after overnight dialysis in reaction buffer in the absence of reducing agent. Lane 5 shows the results of CuP oxidation. Lane 6 shows the results of oxidation followed by the addition of DTT. Lanes 7 and 8 are similar to lane 5 but with increasing concentrations of 3XM. The arrows indicate higher order species (e.g. monomers, dimers, trimers etc.). These data show that under oxidizing conditions (either by dialysis or treatment with CuP), the triple mutant forms ladders (to a much greater extent than the wild type) that can be removed by treatment with the reducing agent DTT. |
![[Figure 4]](yt5033fig4.jpg)
 | STRUCTURAL BIOLOGY |
ISSN: 2059-7983
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