Top-Down Atmospheric Ionization Mass Spectrometry Microscopy Combined With Proteogenomics
| Autor: | Terry Gaasterland, Michael J. Meehan, Pieter C. Dorrestein, Cheng-Chih Hsu, Michael W. Baker, Eduardo R. Macagno |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Tyrosine sulfation Protein mass spectrometry Hypothetical protein Hirudo medicinalis Mass spectrometry Mass Spectrometry Analytical Chemistry 03 medical and health sciences Animals Amino Acid Sequence Protein Processing Proteogenomics Ambient ionization Microscopy Desorption electrospray ionization Chromatography biology Chemistry Neuropeptides Post-Translational Chemical Engineering biology.organism_classification 030104 developmental biology Biochemistry Ganglia Other Chemical Sciences Protein Processing Post-Translational |
| Zdroj: | Analytical chemistry, vol 89, iss 16 Hsu, C-C; Baker, MW; Gaasterland, T; Meehan, MJ; Macagno, ER; & Dorrestein, PC. (2017). Top-Down Atmospheric Ionization Mass Spectrometry Microscopy Combined With Proteogenomics. ANALYTICAL CHEMISTRY, 89(16), 8251-8258. doi: 10.1021/acs.analchem.7b01096. UC San Diego: Retrieved from: http://www.escholarship.org/uc/item/32072890 |
| ISSN: | 1520-6882 0003-2700 |
| Popis: | Mass spectrometry-based protein analysis has become an important methodology for proteogenomic mapping by providing evidence for the existence of proteins predicted at the genomic level. However, screening and identification of proteins directly on tissue samples, where histological information is preserved, remain challenging. Here we demonstrate that the ambient ionization source, nanospray desorption electrospray ionization (nanoDESI), interfaced with light microscopy allows for protein profiling directly on animal tissues at the microscopic scale. Peptide fragments for mass spectrometry analysis were obtained directly on ganglia of the medicinal leech (Hirudo medicinalis) without in-gel digestion. We found that a hypothetical protein, which is predicted by the leech genome, is highly expressed on the specialized neural cells that are uniquely found in adult sex segmental ganglia. Via this top-down analysis, a post-translational modification (PTM) of tyrosine sulfation to this neuropeptide was resolved. This three-in-one platform, including mass spectrometry, microscopy, and genome mining, provides an effective way for mappings of proteomes under the lens of a light microscope. |
| Databáze: | OpenAIRE |
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