title: Supramolecular Affinity Chromatography for Methylation-Targeted Proteomics.
abstract: 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.
authors.affiliation: Department of Chemistry, University of Victoria , Victoria, British Columbia V8W3 V6, Canada.
authors.forename: Graham A E
authors.initials: GA
authors.lastname: Garnett
authors.affiliation: Department of Chemistry, University of Victoria , Victoria, British Columbia V8W3 V6, Canada.
authors.forename: Melissa J
authors.initials: MJ
authors.lastname: Starke
authors.affiliation: Department of Chemistry, University of Victoria , Victoria, British Columbia V8W3 V6, Canada.
authors.forename: Alok
authors.initials: A
authors.lastname: Shaurya
authors.affiliation: Department of Chemistry, University of Victoria , Victoria, British Columbia V8W3 V6, Canada.
authors.forename: Janessa
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Chemistry, University of Victoria , Victoria, British Columbia V8W3 V6, Canada.
authors.forename: Fraser
authors.initials: F
authors.lastname: Hof
mesh_terms: D000818:Animals; D047250:Calixarenes / Q000138:chemical synthesis / Q000737:chemistry; D002417:Cattle; D002846:Chromatography, Affinity; D006657:Histones / Q000032:analysis; D008239:Lysine / Q000031:analogs & derivatives* / Q000032:analysis*; D008745:Methylation; D010455:Peptides / Q000032:analysis*; D040901:Proteomics
chemical_list: D006657:Histones; D010455:Peptides; D047250:Calixarenes; D008239:Lysine