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. Author manuscript; available in PMC: 2020 Nov 1.
Published in final edited form as: Nat Methods. 2020 May 4;17(5):505–508. doi: 10.1038/s41592-020-0821-0

Extended Data Figure 4. Nativeomics defines lipid, peptide and drug bound to the trimeric membrane porin OmpF through progressive dissection using multiple stages of MSn.

Extended Data Figure 4

(a) OmpF bound to lipids is released from detergent micelles (pMS2), the 16+ charge state isolated (red), with lipids released from the complex (pMS3), the peak (m/z 760.5) is then isolated and fragmented to yield fragments at m/z 504.3 and 478.1 (MS4). Spectral matching assigns this lipid as PC 16:0/18:1 (b) OmpF bound to peptide OBS1 is released from detergent micelle (pMS2) and a charge state assigned to the OmpF trimer and three OBS1 peptides was isolated (17+, orange) for pMS3. m/z 777.3 is isolated and fragmented (pMS4) to yield b and y ions that enable peptide sequence determination. (c) OmpF bound to ampicillin. The 17+ charge state (green) was isolated for activation and the ligand released at m/z 350.5 (pMS3). Fragmentation (pMS4) yields characteristic ions that identify ampicillin following database searching or spectral matching. For MS parameters see Online Methods.