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. 2002 Nov;11(11):2575–2583. doi: 10.1110/ps.0220302

Fig. 4.

Fig. 4.

Gel-filtration behavior of wild-type and E206K-K296E double mutant DHODA enzymes under dilute conditions. (A) Wild-type DHODA at 138 μg/mL in the applied sample (black), E206K-K296E double mutant DHODA at 138 μg/mL in the applied sample (red broken line), and E206K-K296E double mutant DHODA at 277 μg/mL in the applied sample (green broken line). The absorbance peak eluting at ∼2350 sec is likely due to small differences between the applied sample and the elution buffer; it did not contain any flavin. (B) E206K-K296E double mutant DHODA at 185 μg/mL in the applied sample (red broken line) and E206K-K296E double mutant DHODA at 185 μg/mL in the applied sample which also contained 600 μM of orotate (green broken line). The part of the elution curve following 2000 sec in B was not shown because of the very strong absorbance from orotate in this region of the profile. The samples were run on a Superose 12 column as described in Materials and Methods.