mitochondria cross-linking mass spectrometry
We performed cross-linking mass spectrometry experiments on intact mitochondria isolated from mouse heart in two conditions, native-state and high-salt treatment to disrupt electrostatic interactions. Both conditions were provided in biological replicates.
Sample Processing Protocol
Mitochondria were directly isolated from mouse heart using centrifugation. Subsequently, they were cross-linking using DSSO cross-linker, followed by standard bottom-up proteomics sample prepration, strong cation exchange (SCX) and LC/MS analysis. Only the later fractions of SCX were analyzed by LC/MS, with MS2 triggering on only speices containing +4 charge and above.
Data Processing Protocol
Data was analyzed using XlinkX v2.0 (Liu et al., Nature Commun, 2017).
Liu F, Lössl P, Rabbitts BM, Balaban RS, Heck AJR. The interactome of intact mitochondria by cross-linking mass spectrometry provides evidence for co-existing respiratory supercomplexes. Mol Cell Proteomics. 2017 PubMed: 29222160