2024
Analysis of Mucin‐Domain Glycoproteins Using Mass Spectrometry
Mahoney K, Malaker S. Analysis of Mucin‐Domain Glycoproteins Using Mass Spectrometry. Current Protocols 2024, 4: e1100. PMID: 38984456, PMCID: PMC11239139, DOI: 10.1002/cpz1.1100.Peer-Reviewed Original Research
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 cellsCheckpointGlycans
2022
A previously uncharacterized O-glycopeptidase from Akkermansia muciniphila requires the Tn-antigen for cleavage of the peptide bond
Medley BJ, Leclaire L, Thompson N, Mahoney KE, Pluvinage B, Parson MAH, Burke JE, Malaker S, Wakarchuk W, Boraston AB. A previously uncharacterized O-glycopeptidase from Akkermansia muciniphila requires the Tn-antigen for cleavage of the peptide bond. Journal Of Biological Chemistry 2022, 298: 102439. PMID: 36049519, PMCID: PMC9513282, DOI: 10.1016/j.jbc.2022.102439.Peer-Reviewed Original ResearchConceptsUncharacterized proteinsCarbohydrate-binding moduleHuman gut microbiotaHost adaptationSubstrate recognitionCatalytic motifN-terminalHost healthGut environmentPeptidase activityHost mucinsNutrient sourceAkkermansia muciniphilaEnzymeKey membersPeptide bondProteinBacteriumActive siteGut microbiotaTn antigenPeptide backboneGenesMicrobesCurrent 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 developmentsRevealing the human mucinome
Malaker SA, Riley NM, Shon DJ, Pedram K, Krishnan V, Dorigo O, Bertozzi CR. Revealing the human mucinome. Nature Communications 2022, 13: 3542. PMID: 35725833, PMCID: PMC9209528, DOI: 10.1038/s41467-022-31062-4.Peer-Reviewed Original ResearchConceptsModular protein domainsKey molecular signaturesProtein domainsTransmembrane proteinDetection of hundredsMucin domainGlycoprotein identificationHuman diseasesMolecular signaturesCell lysatesEnrichment strategyGlycoproteinVivo sourceOverlapping populationsImportant roleDomainStcEProteinLysatesOvarian cancer patientsComplex samples
2021
Mucinomics as the Next Frontier of Mass Spectrometry
Rangel-Angarita V, Malaker SA. Mucinomics as the Next Frontier of Mass Spectrometry. ACS Chemical Biology 2021, 16: 1866-1883. PMID: 34319686, DOI: 10.1021/acschembio.1c00384.Peer-Reviewed Original ResearchClassification, structural biology, and applications of mucin domain-targeting proteases
Shon DJ, Kuo A, Ferracane MJ, Malaker SA. Classification, structural biology, and applications of mucin domain-targeting proteases. Biochemical Journal 2021, 478: 1585-1603. PMID: 33909028, DOI: 10.1042/bcj20200607.Peer-Reviewed Original ResearchConceptsClass of proteinsHost-microbe interfaceImmunoglobulin A (IgA) proteaseThreonine residuesGlycan repertoireStructural biologyDistinct bacteriaGlycan structuresSource of nutrientsHinge regionProteaseIgA proteaseIntricate interplayMucosal environmentColonizationStructural featuresMutualistsMucosal colonizationEpithelial surfaceBacterial colonizationMicrobesTranslocation of microbesBenefits of Chemical Sugar Modifications Introduced by Click Chemistry for Glycoproteomic Analyses
Calle B, Bineva-Todd G, Marchesi A, Flynn H, Ghirardello M, Tastan OY, Roustan C, Choi J, Galan MC, Schumann B, Malaker SA. Benefits of Chemical Sugar Modifications Introduced by Click Chemistry for Glycoproteomic Analyses. Journal Of The American Society For Mass Spectrometry 2021, 32: 2366-2375. PMID: 33871988, PMCID: PMC7611619, DOI: 10.1021/jasms.1c00084.Peer-Reviewed Original ResearchConceptsMass spectrometryCharge densityMass spectrometric signatureLow charge densityIntact O-glycopeptidesHigh charge densityTandem mass spectrometryClick chemistryChemical methodsChemical modificationO-glycopeptidesETD fragmentationFragmentation behaviorMonosaccharide analoguesSpectrometric signaturesMucin-type O-glycansGlycoproteomic analysisCharge stateComplex post-translational modificationsRight analytical toolsGlycan structuresO-glycosylationSugar modificationsGlycopeptidesSpectrometry
2020
An enzymatic toolkit for selective proteolysis, detection, and visualization of mucin-domain glycoproteins
Shon DJ, Malaker SA, Pedram K, Yang E, Krishnan V, Dorigo O, Bertozzi CR. An enzymatic toolkit for selective proteolysis, detection, and visualization of mucin-domain glycoproteins. Proceedings Of The National Academy Of Sciences Of The United States Of America 2020, 117: 21299-21307. PMID: 32817557, PMCID: PMC7474620, DOI: 10.1073/pnas.2012196117.Peer-Reviewed Original Research
2019
The mucin-selective protease StcE enables molecular and functional analysis of human cancer-associated mucins
Malaker SA, Pedram K, Ferracane MJ, Bensing BA, Krishnan V, Pett C, Yu J, Woods EC, Kramer JR, Westerlind U, Dorigo O, Bertozzi CR. The mucin-selective protease StcE enables molecular and functional analysis of human cancer-associated mucins. Proceedings Of The National Academy Of Sciences Of The United States Of America 2019, 116: 7278-7287. PMID: 30910957, PMCID: PMC6462054, DOI: 10.1073/pnas.1813020116.Peer-Reviewed Original ResearchConceptsModular protein domainsProtein domainsSecreted proteinsMucin domainFunctional analysisHuman diseasesMucin biologyGlycosylation patternsCultured cellsMolecular levelCell surfaceSequence coverageKey playersBacterial proteasesCancer-associated mucinsDiscrete peptidesBiological ligandsProteaseHuman mucinsGlycoform analysisStcESiglec-9Domain structureMass spectrometryDomain