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. 2021 May 12;11:10143. doi: 10.1038/s41598-021-89575-9

Figure 1.

Figure 1

Purification and subunit composition of P. denitrificans complex I. (a) Typical Ni-affinity chromatography trace produced using a HisTrap HP column. The column was washed with 80 mM imidazole and protein eluted with 200 mM imidazole. (b) Typical size exclusion chromatography trace using a Superdex 200 increase 5/150 GL column. Complex I was identified by measuring the relative NADH:APAD+ activity in eluted fractions (black trace). (c) SDS-PAGE analysis of purified P. denitrificans complex I. Two full-length lanes are shown from a single gel; the image has been cut and they have been moved to be adjacent to each other. The original image is shown in SI Fig. S6. Individual complex I subunits were identified and assigned by excising each band, treating the sample with trypsin and analyzing the resultant peptides by mass spectrometry. Peptides were assigned to a subunit/protein by peptide mass fingerprinting.