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Figure 2
DUOX and examples of electron and neutron density maps for the DUOX–8AZA–W1 complex. (a) SDS-PAGE analysis of purified DUOX and HUOX (protiated, for comparison) samples. Protein markers are on the left-hand side lane with their molecular weights indicated. (b), (c) ESI-MS analysis of purified (b) DUOX and (c) HUOX, showing a mass difference of 1803 a.m.u. As the calculated mass difference (assuming complete deuteration) is 1820 a.m.u., this corresponds to ∼99.1% deuterium incorporation in the DUOX sample. (d), (e) Representative examples of 2mFo − DFc (d) electron density and (e) neutron maps contoured at the 1.0σ level. X-ray and neutron data extend to 1.33 and 2.10 Å, respectively. As in Fig. 1[link](c), residues shown in yellow and magenta belong to different UOX protomers. Nitro­gen, oxygen and deuterium atoms are shown in blue, red and wheat, respectively. The use of perdeuterated UOX provides neutron maps essentially free from cancellation effects affording clear visualization of both non-exchangeable and exchangeable hydrogens.

IUCrJ
Volume 8| Part 1| January 2021| Pages 46-59
ISSN: 2052-2525
https://doi.org/10.1107/S2052252520013615