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Reassembling protein complexes after controlled disassembly by top-down mass spectrometry in native mode

ABSTRACT: The native top-down mass spectrometry (nTDMS) platform is being applied in structural biology and discovery proteomics with great success, as it provides three levels of information in a single experiment: the intact mass of a protein or complex, the masses of its subunits and non-covalent cofactors, and subunit fragment masses that capture the primary sequence and combinations of diverse post-translational modifications (PTMs). While the intact mass data are readily converted deconvoluted using well-known software options, the analysis of fragmentation data that result from a tandem MS experiment, essential for proteoform characterization, is not yet standardized. In this tutorial, we offer a decision-tree for the analysis of subunit fragmentation data that result from nTDMS experiments on protein complexes and on diverse bioassemblies. We include an overview of strategies for navigating this type of analysis, provide example data sets, and highlight software for the hypothesis-driven interrogation of fragment ions for localization of PTMs, metals, and cofactors on native proteoforms.

INSTRUMENT(S): Q Exactive HF EMR

ORGANISM(S): Bos Taurus (ncbitaxon:9913) Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Neil Kelleher

PROVIDER: MSV000086404 | MassIVE | Tue Nov 03 11:00:00 GMT 2020

REPOSITORIES: MassIVE

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Reassembling protein complexes after controlled disassembly by top-down mass spectrometry in native mode

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