Combining Surface-Induced Dissociation and Charge Detection Mass Spectrometry to Reveal the Native Topology of Heterogeneous Protein Complexes.
Chen DuSean P ClearyMarius M KostelicBenjamin J JonesJared O KafaderVicki H WysockiPublished in: Analytical chemistry (2023)
Charge detection mass spectrometry (CDMS) enables the direct mass measurement of heterogeneous samples on the megadalton scale, as the charge state for a single ion is determined simultaneously with the mass-to-charge ratio ( m / z ). Surface-induced dissociation (SID) is an effective activation method to dissociate non-intertwined, non-covalent protein complexes without extensive gas-phase restructuring, producing various subcomplexes reflective of the native protein topology. Here, we demonstrate that using CDMS after SID on an Orbitrap platform offers subunit connectivity, topology, proteoform information, and relative interfacial strengths of the intact macromolecular assemblies. SID dissects the capsids (∼3.7 MDa) of adeno-associated viruses (AAVs) into trimer-containing fragments (3mer, 6mer, 9mer, 15mer, etc.) that can be detected by the individual ion mass spectrometry (I2MS) approach on Orbitrap instruments. SID coupled to CDMS provides unique structural insights into heterogeneous assemblies that are not readily obtained by traditional MS measurements.
Keyphrases
- mass spectrometry
- liquid chromatography
- gas chromatography
- high resolution
- capillary electrophoresis
- high performance liquid chromatography
- high glucose
- high resolution mass spectrometry
- protein protein
- diabetic rats
- tandem mass spectrometry
- amino acid
- real time pcr
- healthcare
- label free
- loop mediated isothermal amplification
- drug induced
- small molecule
- gene expression
- signaling pathway
- oxidative stress
- cell proliferation
- health information
- quantum dots
- ionic liquid
- ultra high performance liquid chromatography
- cell death
- breast cancer cells
- patient reported outcomes