Back to Search
Start Over
Enhancing Top-Down Proteomics of Brain Tissue with FAIMS
Enhancing Top-Down Proteomics of Brain Tissue with FAIMS
- Source :
- J Proteome Res
- Publication Year :
- 2021
- Publisher :
- American Chemical Society (ACS), 2021.
-
Abstract
- Proteomic investigations of Alzheimer’s and Parkinson’s disease have provided valuable insights into neurodegenerative disorders. Thus far, these investigations have largely been restricted to bottom-up approaches, hindering the degree to which one can characterize a protein’s “intact” state. Top-down proteomics (TDP) overcomes this limitation; however, it is typically limited to observing only the most abundant proteoforms and of a relatively small size. Therefore, fractionation techniques are commonly used to reduce sample complexity. Here, we investigate gas-phase fractionation through high-field asymmetric waveform ion mobility spectrometry (FAIMS) within TDP. Utilizing a high complexity sample derived from Alzheimer’s disease (AD) brain tissue, we describe how the addition of FAIMS to TDP can robustly improve the depth of proteome coverage. For example, implementation of FAIMS with external compensation voltage (CV) stepping at −50, −40, and −30 CV could more than double the mean number of non-redundant proteoforms, genes, and proteome sequence coverage compared to without FAIMS. We also found that FAIMS can influence the transmission of proteoforms and their charge envelopes based on their size. Importantly, FAIMS enabled the identification of intact amyloid beta (Aβ) proteoforms, including the aggregation-prone Aβ(1–42) variant which is strongly linked to AD. Raw data and associated files have been deposited to the ProteomeXchange Consortium via the MassIVE data repository with data set identifier PXD023607.
- Subjects :
- Proteomics
0301 basic medicine
Proteome
Ion-mobility spectrometry
Amyloid beta
Brain tissue
Computational biology
Top-down proteomics
Biochemistry
Article
Gas phase
03 medical and health sciences
Ion Mobility Spectrometry
Brain Chemistry
Amyloid beta-Peptides
030102 biochemistry & molecular biology
biology
Sample complexity
Chemistry
Brain
General Chemistry
030104 developmental biology
biology.protein
Asymmetric waveform
Subjects
Details
- ISSN :
- 15353907 and 15353893
- Volume :
- 20
- Database :
- OpenAIRE
- Journal :
- Journal of Proteome Research
- Accession number :
- edsair.doi.dedup.....f8c4bbadfb090cd5eca76d44e3c96d14
- Full Text :
- https://doi.org/10.1021/acs.jproteome.1c00049