Proteoform-selective imaging of tissues using mass spectrometry

21 January 2022, Version 2
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

Unraveling the complexity of biological systems relies on the development of new approaches for spatially resolved proteoform-specific analysis of the proteome. Herein, we employ nanospray desorption electrospray ionization mass spectrometry imaging (nano-DESI MSI) for the proteoform-selective imaging of biological tissues. Nano-DESI generates multiply charged protein ions, which is advantageous for their structural characterization using tandem mass spectrometry (MS/MS) directly on the tissue. Proof-of-concept experiments demonstrate that nano-DESI MSI combined with on-tissue top-down proteomics is ideally suited for the proteoform-selective imaging of tissue sections. Using rat brain tissue as a model system, we provide the first evidence of differential proteoform expression in different regions of the brain.

Keywords

proteoforms
post-translational modifications
on-tissue top-down proteomics
mass spectrometry imaging

Supplementary materials

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Description
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Supporting information
Description
This document contains the descriptions of methods and experiments, and supplemental figures.
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Supporting table
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This document contains the proteoforms assignments.
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