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Figure 1
Biochemical characterization of NmqNOR. (a) Size-exclusion chromatography of NmqNOR. The solid curve shows the BRIL construct of NmqNOR and the broken curve shows the wild-type NmqNOR trace at 280 nm absorbance. Two types of fractions were observed in both constructs, which are indicated by the labels P1 for dimer and P2 for monomer (the red dashed line shows the divide between peak 1 and peak 2). (b) Blue native gel experiment for both fractions (P1 and P2) after concentration in 100 kDa cutoff centrifugal filtration devices (Amicon). Both 5 µg (labelled 5) and 10 µg (labelled 10) of each fraction was loaded onto the gel. Peak 1 samples migrate as the higher molecular-weight species (∼242 kDa), whilst peak 2 samples migrate between 66 and 146 kDa. Two bands of the high and low molecular-weight species could be observed in both peak fractions when loaded with 10 µg sample, which indicate a slight mixture of monomer and dimer in both peak 1 and peak 2. Lane M, native PAGE marker, with values corresponding to molecular weight in kDa. (c) NO-reduction activity measurement of both peak 1 and peak 2 fractions. Peak 1 (solid curve) shows a higher NO-reduction activity compared with peak 2 (broken curve).

IUCrJ
Volume 7| Part 3| May 2020| Pages 404-415
ISSN: 2052-2525
https://doi.org/10.1107/S2052252520003656