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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
Published on: November 15, 2017
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Top-Down Atmospheric Ionization Mass Spectrometry Microscopy Combined With Proteogenomics
Cheng-Chih Hsu1, Michael W Baker, Terry Gaasterland
1Department of Chemistry, National Taiwan University , Taipei 10617, Taiwan.
Analytical Chemistry
|July 11, 2017
Summary
Mass spectrometry now profiles proteins on intact animal tissues using nanospray desorption electrospray ionization (nanoDESI) microscopy. This method identified a novel neuropeptide and its tyrosine sulfation modification in leech neural cells.
Area of Science:
- Proteomics
- Molecular Biology
- Neuroscience
Background:
- Mass spectrometry is crucial for proteogenomic mapping, confirming genomic predictions.
- Direct protein analysis on tissue sections with preserved histology is challenging.
Purpose of the Study:
- To develop a method for direct, microscopic-scale protein profiling on animal tissues.
- To identify and characterize proteins and their modifications in situ.
Main Methods:
- Utilized nanospray desorption electrospray ionization (nanoDESI) coupled with light microscopy.
- Performed direct peptide analysis on medicinal leech ganglia without in-gel digestion.
- Integrated mass spectrometry, microscopy, and genome mining.
Main Results:
- Achieved direct protein profiling on leech ganglia at the microscopic level.
- Identified a hypothetical neuropeptide highly expressed in specialized neural cells.
- Resolved a post-translational modification (tyrosine sulfation) on the identified neuropeptide.
Conclusions:
- The nanoDESI-microscopy platform enables in situ proteome mapping on tissues.
- This approach facilitates the discovery of novel proteins and post-translational modifications in their native context.
- Combines imaging, mass spectrometry, and genomics for comprehensive proteomic analysis.
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