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Cell-Type Specific Protein Purification and Identification from Complex Tissues Using a Mutant Methionine tRNA Synthetase Mouse Line
Published on: April 13, 2022
Mass spectrometric analysis of mutant mice
Simon J North1, Jihye Jang-Lee, Rebecca Harrison
1Division of Molecular Biosciences, Faculty of Natural Sciences, Imperial College London, London, United Kingdom.
Methods in Enzymology
|September 7, 2010
Summary
Mass spectrometry (MS) is a powerful tool for glycan analysis in biological samples. This study details MS methods for mouse tissues, aiding in understanding glycosylation changes in knockout models.
Area of Science:
- Glycomics
- Mass Spectrometry
- Mouse Models
Background:
- Mass spectrometry (MS) is crucial for analyzing glycan structures in various biological sources.
- Matrix-assisted laser desorption ionization time of flight (MALDI-TOF) and complementary techniques generate glycomic mass fingerprints.
- Mice are valuable model organisms for studying human biology and disease due to genetic homology and targeted mutagenesis capabilities.
Purpose of the Study:
- To detail methodologies for glycan pool preparation, derivatization, purification, and MS analysis from mouse organs and tissues.
- To present a summary of glycomic data from wild-type and glycosylation-knockout mouse tissues.
- To provide a detailed example of glycomic analysis for the Mgat4a knockout mouse.
Main Methods:
- Utilizing mass spectrometry (MS), including MALDI-TOF and tandem MS (MS/MS) with electrospray (ES).
- Employing complementary techniques such as chemical/enzymatic digestions and GC-MS linkage analysis.
- Applying bioinformatic tools for detailed structural elucidation of glycans.
Main Results:
- Established MS-based glycomic analysis protocols for mouse tissues.
- Generated a repository of structural data for wild-type and knockout mouse tissues through the Consortium for Functional Glycomics (CFG).
- Demonstrated the identification of glycomic perturbations in genetically modified mice.
Conclusions:
- MS-based glycomic analysis of mouse models provides critical insights into glycosylation.
- The described methodologies and data repository facilitate glycobiology research.
- Comparative glycomic analysis of knockout and wild-type mice aids in understanding gene function in glycosylation.
