Supramolecular Affinity Chromatography for Methylation-Targeted Proteomics
| Autor: | Alok Shaurya, Melissa J. Starke, Fraser Hof, Graham A. E. Garnett, Janessa Li |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
Proteomics
0301 basic medicine Supramolecular chemistry Context (language use) 010402 general chemistry Mass spectrometry Methylation 01 natural sciences Chromatography Affinity Analytical Chemistry Histones 03 medical and health sciences Methyllysine chemistry.chemical_compound Affinity chromatography Animals Chromatography biology Chemistry Lysine 0104 chemical sciences 030104 developmental biology Polyclonal antibodies biology.protein Cattle Calixarenes Peptides |
| Zdroj: | Analytical Chemistry. 88:3697-3703 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.5b04508 |
| Popis: | Proteome-wide studies of post-translationally methylated species using mass spectrometry are complicated by high sample diversity, competition for ionization among peptides, and mass redundancies. Antibody-based enrichment has powered methylation proteomics until now, but the reliability, pan-specificity, polyclonal nature, and stability of the available pan-specific antibodies are problematic and do not provide a standard, reliable platform for investigators. We have invented an anionic supramolecular host that can form host-guest complexes selectively with methyllysine-containing peptides and used it to create a methylysine-affinity column. The column resolves peptides on the basis of methylation-a feat impossible with a comparable commercial cation-exchange column. A proteolyzed nuclear extract was separated on the methyl-affinity column prior to standard proteomics analysis. This experiment demonstrates that such chemical methyl-affinity columns are capable of enriching and improving the analysis of methyllysine residues from complex protein mixtures. We discuss the importance of this advance in the context of biomolecule-driven enrichment methods. |
| Databáze: | OpenAIRE |
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