Fig. 1: High-resolution complexome profiling of mitochondria using improved csBN–MS.

a, Schematic of the csBN–MS workflow, comprising protein separation by blue native gel electrophoresis (BN-PAGE), cryo-slicing of the gel, MS analysis, data processing and protein quantification. The interactive online platform presents all resulting abundance–mass profiles and MS data for user-directed inspection and evaluation. LC–MS/MS, liquid chromatography coupled with tandem MS. b, Abundance profiles over slice number and apparent molecular mass for the indicated set of mitochondrial proteins; the profiles were normalized to the maximal abundance value determined for the respective protein. c, Map of protein peaks determined from abundance–mass profiles by multi-Gaussian fits and verified by manual inspection. All of the symbols represent identified peak parameters (apparent molecular mass, abundance and half-width as indicated at the top right). The shaded areas denote peaks with maxima outside the blue native gel. Coloured frames denote known protein complexes that are assembled from the indicated constituents of which the peaks are highlighted in the same colour. Insets: sections of abundance–mass profiles determined for the indicated protein constituents of established mitochondrial assemblies together with the fit lines (dashed) obtained by Gaussian fitting. For Phb1 and Phb2, the composite fit lines (solid, blue) and the individual components (dashed, grey, black) are shown. Note the close match in the midpoint of the individual Gaussian functions obtained in independent fits for the constituents of the same complex.