2024
Mass Spectrometry-Compatible Elution Technique Enables an Improved Mucin-Selective Enrichment Strategy to Probe the Mucinome
Mahoney K, Chang V, Lucas T, Maruszko K, Malaker S. Mass Spectrometry-Compatible Elution Technique Enables an Improved Mucin-Selective Enrichment Strategy to Probe the Mucinome. Analytical Chemistry 2024, 96: 5242-5250. PMID: 38512228, DOI: 10.1021/acs.analchem.3c05762.Peer-Reviewed Original ResearchMucin-domain glycoproteinsO-glycopeptidesGlycopeptide signalsIn-gel digestionMass spectrometryElution conditionsSolid supportBinding moietyElution stepDownstream analysisO-glycosylationIn-gelBiological functionsElutionHuman serumEnrichment strategyCell linesMoietyEffective isolationSpectrometryCatalyticallyGlycoproteinSampling requirementsGlycopeptidesStcE
2023
Glycoproteomic landscape and structural dynamics of TIM family immune checkpoints enabled by mucinase SmE
Chongsaritsinsuk J, Steigmeyer A, Mahoney K, Rosenfeld M, Lucas T, Smith C, Li A, Ince D, Kearns F, Battison A, Hollenhorst M, Judy Shon D, Tiemeyer K, Attah V, Kwon C, Bertozzi C, Ferracane M, Lemmon M, Amaro R, Malaker S. Glycoproteomic landscape and structural dynamics of TIM family immune checkpoints enabled by mucinase SmE. Nature Communications 2023, 14: 6169. PMID: 37794035, PMCID: PMC10550946, DOI: 10.1038/s41467-023-41756-y.Peer-Reviewed Original ResearchConceptsFamily of proteinsMucin domainO-glycosylationBiological functionsKey regulatorComplex glycansMass spectrometric analysisFunctional relevanceTIM familyDetailed molecular structureCritical roleGlycosylationProteinSpectrometric analysisStructural featuresUnique abilityStructural dynamicsMolecular dynamics simulationsTim-3 functionFamilyPowerful workflowRegulatorImmune cellsCheckpointGlycansBump-and-hole engineering of human polypeptide N-acetylgalactosamine transferases to dissect their protein substrates and glycosylation sites in cells
Calle B, Gonzalez-Rodriguez E, Mahoney K, Cioce A, Bineva-Todd G, Tastan O, Roustan C, Flynn H, Malaker S, Schumann B. Bump-and-hole engineering of human polypeptide N-acetylgalactosamine transferases to dissect their protein substrates and glycosylation sites in cells. STAR Protocols 2023, 4: 101974. PMID: 36633947, PMCID: PMC9843269, DOI: 10.1016/j.xpro.2022.101974.Peer-Reviewed Original ResearchConceptsProtein substratesGlycosylation sitesGalNAc-T familyTransfection of cellsIndividual glycosyltransferasesBioorthogonal reportersN-acetylgalactosamine transferaseSubstrate specificityBiological functionsGalNAc-TsDisease relevanceMolecular biologyComplete detailsGlycosyltransferasesTransferaseCellsBiosynthesisBiologyReporterTransfectionGlycansSubstrateEnzymeHole engineeringSites
2022
Current strategies for characterization of mucin-domain glycoproteins
Ince D, Lucas TM, Malaker SA. Current strategies for characterization of mucin-domain glycoproteins. Current Opinion In Chemical Biology 2022, 69: 102174. PMID: 35752002, DOI: 10.1016/j.cbpa.2022.102174.Peer-Reviewed Original ResearchConceptsGlycopeptide mimeticsPost-translational modificationsCurrent characterization techniquesCellular glycosylation pathwaysSynthetic methodCharacterization techniquesGlycosylation pathwayMucin domainO-glycosylationBiological functionsGlycoproteomic workflowGlycosylationGlycoproteinExciting avenuesRecent breakthroughsMucin glycoproteinsRecent developments