2024
Quantification and Site-Specific Analysis of Co-occupied N- and O‑Glycopeptides
Chongsaritsinsuk J, Rangel-Angarita V, Lucas T, Mahoney K, Enny O, Katemauswa M, Malaker S. Quantification and Site-Specific Analysis of Co-occupied N- and O‑Glycopeptides. Journal Of Proteome Research 2024 PMID: 39498894, DOI: 10.1021/acs.jproteome.4c00574.Peer-Reviewed Original ResearchElectron-based methodsO-glycopeptidesElectron-based fragmentation methodsFragmentation methodCo-occupancyO-glycosylated peptidesAnalysis of glycopeptidesPost-translational modificationsN-glycoproteomic analysisCore 1 structureComplex samplesO-glycansPurified proteinProtein glycosylationO-glycositesN-glycoproteomeNonmucin glycoproteinsTryptic peptidesDissociation methodO-linkedAnalysis of mucinsSite-specific analysisGlycoproteinGlycosylationProtein
2022
Cell-specific bioorthogonal tagging of glycoproteins
Cioce A, Calle B, Rizou T, Lowery SC, Bridgeman VL, Mahoney KE, Marchesi A, Bineva-Todd G, Flynn H, Li Z, Tastan OY, Roustan C, Soro-Barrio P, Rafiee MR, Garza-Garcia A, Antonopoulos A, Wood TM, Keenan T, Both P, Huang K, Parmeggian F, Snijders AP, Skehel M, Kjær S, Fascione MA, Bertozzi CR, Haslam SM, Flitsch SL, Malaker SA, Malanchi I, Schumann B. Cell-specific bioorthogonal tagging of glycoproteins. Nature Communications 2022, 13: 6237. PMID: 36284108, PMCID: PMC9596482, DOI: 10.1038/s41467-022-33854-0.Peer-Reviewed Original ResearchConceptsMass spectrometry glycoproteomicsArtificial biosynthetic pathwayTumor-derived cell linesCellular model systemNon-transfected cellsCellular functionsProtein glycosylationBiosynthetic pathwayProteome analysisGlycosylation sitesBioorthogonal tagsCancer developmentCell linesModel systemImportant modulatorIntricate interactionsCo-culture modelGlycoproteinCellsGlycoprotein expressionMouse modelGlycoproteomeGlycosylationTaggingMonocultureGlycoproteomics
Bagdonaite I, Malaker S, Polasky D, Riley N, Schjoldager K, Vakhrushev S, Halim A, Aoki-Kinoshita K, Nesvizhskii A, Bertozzi C, Wandall H, Parker B, Thaysen-Andersen M, Scott N. Glycoproteomics. Nature Reviews Methods Primers 2022, 2: 48. DOI: 10.1038/s43586-022-00128-4.Peer-Reviewed Original ResearchGlycan structuresMass spectrometryPost-translational additionIntact glycopeptide analysisSite of modificationProtein glycosylationProtein modificationBioinformatics platformBiological processesGlycopeptide analysisMS fragmentationGlycoproteomic methodsGlycoproteomicsGlycosylationProtein isolationProteolytic digestionPeptide sequencesSystem-wide contextStudy of glycopeptidesPrimersRecent advancesExciting fieldProteinGlycansSpectrometry
2020
Generating orthogonal glycosyltransferase and nucleotide sugar pairs as next-generation glycobiology tools
Cioce A, Malaker SA, Schumann B. Generating orthogonal glycosyltransferase and nucleotide sugar pairs as next-generation glycobiology tools. Current Opinion In Chemical Biology 2020, 60: 66-78. PMID: 33125942, PMCID: PMC7955280, DOI: 10.1016/j.cbpa.2020.09.001.Peer-Reviewed Original ResearchConceptsMass spectrometry glycoproteomicsCell surface glycoproteomeProtein glycosylationBiological processesQuantitative biologyMutant glycosyltransferasesGlycoproteomicsDifferent glycansGlycansPrecision toolsGlycosyltransferasesBiosynthesisGlycoproteomeGlycomeGlycosyltransferaseBiologyGlycosylationGlycobiologyLabeling reagentLimited specificityPhysiologySpecificityParticular subtypeRoleCellsMetabolic precision labeling enables selective probing of O-linked N-acetylgalactosamine glycosylation
Debets MF, Tastan OY, Wisnovsky SP, Malaker SA, Angelis N, Moeckl LKR, Choi J, Flynn H, Wagner LJS, Bineva-Todd G, Antonopoulos A, Cioce A, Browne WM, Li Z, Briggs DC, Douglas HL, Hess GT, Agbay AJ, Roustan C, Kjaer S, Haslam SM, Snijders AP, Bassik MC, Moerner WE, Li VSW, Bertozzi CR, Schumann B. Metabolic precision labeling enables selective probing of O-linked N-acetylgalactosamine glycosylation. Proceedings Of The National Academy Of Sciences Of The United States Of America 2020, 117: 25293-25301. PMID: 32989128, PMCID: PMC7568240, DOI: 10.1073/pnas.2007297117.Peer-Reviewed Original ResearchConceptsStructure-based design processN-acetylgalactosamine glycosylationProtein glycosylation eventsCell surface glycoproteomeCRISPR knockout screensMetabolic labeling experimentsGlycan subtypesGlycosylation eventsSecretory pathwayGalNAc glycosylationProtein glycosylationEctopic expressionSer/GalNAc-T2Superresolution microscopyGlycan typesLiving cellsBioorthogonal tagsGlycosylationIntestinal organoidsMetabolic interconversionSuch specificityUridine diphosphateLabeling experimentsCellsBump-and-Hole Engineering Identifies Specific Substrates of Glycosyltransferases in Living Cells
Schumann B, Malaker SA, Wisnovsky SP, Debets MF, Agbay AJ, Fernandez D, Wagner LJS, Lin L, Li Z, Choi J, Fox DM, Peh J, Gray MA, Pedram K, Kohler JJ, Mrksich M, Bertozzi CR. Bump-and-Hole Engineering Identifies Specific Substrates of Glycosyltransferases in Living Cells. Molecular Cell 2020, 78: 824-834.e15. PMID: 32325029, PMCID: PMC7276986, DOI: 10.1016/j.molcel.2020.03.030.Peer-Reviewed Original ResearchConceptsLiving cellsPolypeptide N-acetylgalactosaminyl transferasesCell surface glycomesEssential biological processesComplex biosynthetic machineryMajor physiological processesN-acetylgalactosaminyl transferaseBiosynthetic regulationBiosynthetic machineryGlycosyltransferase familyIndividual glycosyltransferasesProtein glycosylationPosttranslational modificationsGlycan fine structureBiosynthetic pathwayPhysiological contextBiological processesPhysiological processesGlycan structuresSpecific substratesGlycosyltransferasesChemical functionalityExperimental strategiesCellsBiosynthesis