Combining native and ‘omics’ mass spectrometry to identify endogenous ligands bound to membrane proteins

Ligands bound to protein assemblies provide critical information for function, yet are often difficult to capture and define. Here we develop a top-down method, ‘nativeomics’, unifying ‘omics’ (lipidomics, proteomics, metabolomics) analysis with native mass spectrometry to identify ligands bound to membrane protein assemblies. By maintaining the link between proteins and ligands, we define the lipidome/metabolome in contact with membrane porins and a mitochondrial translocator to discover potential regulators of protein function. ‘Nativeomics’ enables identification of ligands bound to membrane proteins through detection of intact protein–ligand assemblies followed by dissociation and identification of individual ligands within the same mass spectrometry experiment.

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