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 ResearchGlycoproteomics: Charting new territory in mass spectrometry and glycobiology
Malaker S. Glycoproteomics: Charting new territory in mass spectrometry and glycobiology. Journal Of Mass Spectrometry 2024, 59: e5034. PMID: 38726698, DOI: 10.1002/jms.5034.Peer-Reviewed Original Research
2023
Mutational screens highlight glycosylation as a modulator of colony-stimulating factor 3 receptor (CSF3R) activity
Hollander M, Malaker S, Riley N, Perez I, Abney N, Gray M, Maxson J, Cochran J, Bertozzi C. Mutational screens highlight glycosylation as a modulator of colony-stimulating factor 3 receptor (CSF3R) activity. Journal Of Biological Chemistry 2023, 299: 104755. PMID: 37116708, PMCID: PMC10245049, DOI: 10.1016/j.jbc.2023.104755.Peer-Reviewed Original ResearchConceptsProtein domain mappingSingle amino acid mutationLigand-independent activityChronic neutrophilic leukemiaCell surface receptorsAmino acid mutationsColony-stimulating factor 3 receptorSerine residuesPresence of GalNAcHotspot residuesNeutrophilic leukemiaHuman diseasesAcid mutationsReceptor signalingAmino acidsCell growthSurface receptorsReceptor alpha chainMutationsAbundant typeDomain mappingDisease mechanismsUncontrolled activityAlpha chainWhite blood cellsVibrio cholerae biofilms use modular adhesins with glycan-targeting and nonspecific surface binding domains for colonization
Huang X, Nero T, Weerasekera R, Matej K, Hinbest A, Jiang Z, Lee R, Wu L, Chak C, Nijjer J, Gibaldi I, Yang H, Gamble N, Ng W, Malaker S, Sumigray K, Olson R, Yan J. Vibrio cholerae biofilms use modular adhesins with glycan-targeting and nonspecific surface binding domains for colonization. Nature Communications 2023, 14: 2104. PMID: 37055389, PMCID: PMC10102183, DOI: 10.1038/s41467-023-37660-0.Peer-Reviewed Original ResearchConceptsBiofilm matrix exopolysaccharideFacilitate host colonizationVibrio cholerae biofilmsΒ-propeller domainMatrix exopolysaccharideModular domainsHost colonizationRedundant rolesDistinct functionsAbiotic surfacesAdhesive proteinsHost surfaceHuman pathogensVibrio choleraeAdhesinsBacterial biofilmsHost tissuesColonization modelColonizationBAP1BiofilmsPathogensAntibiotic resistanceRBMCDomainComprehensive analysis of platelet glycoprotein Ibα ectodomain glycosylation
Hollenhorst M, Tiemeyer K, Mahoney K, Aoki K, Ishihara M, Lowery S, Rangel-Angarita V, Bertozzi C, Malaker S. Comprehensive analysis of platelet glycoprotein Ibα ectodomain glycosylation. Journal Of Thrombosis And Haemostasis 2023, 21: 995-1009. PMID: 36740532, PMCID: PMC10065957, DOI: 10.1016/j.jtha.2023.01.009.Peer-Reviewed Original ResearchConceptsO-glycositesGPIb-IX-V complexMajor ligand-binding subunitAmino acid sitesLigand-binding subunitVon Willebrand factor bindingPlatelet glycoprotein IbαMechanosensory domainFactor bindingEndogenous proteinsRecombinant proteinsN-glycositesStructural rolePlatelet biologyGlycan structuresGlycoprotein IbαO-glycansT antigenGlycosylation profileDiverse repertoireGlycosylationGlycansComprehensive analysisGlycositesVon Willebrand factorBump-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
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 backboneGenesMicrobesRevealing 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 samplesMucus sialylation determines intestinal host-commensal homeostasis
Yao Y, Kim G, Shafer S, Chen Z, Kubo S, Ji Y, Luo J, Yang W, Perner SP, Kanellopoulou C, Park AY, Jiang P, Li J, Baris S, Aydiner EK, Ertem D, Mulder DJ, Warner N, Griffiths AM, Topf-Olivestone C, Kori M, Werner L, Ouahed J, Field M, Liu C, Schwarz B, Bosio CM, Ganesan S, Song J, Urlaub H, Oellerich T, Malaker SA, Zheng L, Bertozzi CR, Zhang Y, Matthews H, Montgomery W, Shih HY, Jiang J, Jones M, Baras A, Shuldiner A, Gonzaga-Jauregui C, Snapper SB, Muise AM, Shouval DS, Ozen A, Pan KT, Wu C, Lenardo MJ. Mucus sialylation determines intestinal host-commensal homeostasis. Cell 2022, 185: 1172-1188.e28. PMID: 35303419, PMCID: PMC9088855, DOI: 10.1016/j.cell.2022.02.013.Peer-Reviewed Original ResearchConceptsInflammatory bowel diseaseMicrobial pathogen-associated molecular patternsPathogen-associated molecular patternsMicrobial symbiosisRegulatory networksIntestinal mucusGlycoproteomic profilingMolecular patternsTerminal sialylationMucus proteinsProteolytic degradationBiochemical analysisBacterial invasionBowel diseaseIntestinal inflammationMucus integrityMutationsSialylationST6GALNAC1Barrier integrityGoblet cellsMucus barrierFirst lineSymbiosisCommensalism
2021
Interleukin-22 regulates B3GNT7 expression to induce fucosylation of glycoproteins in intestinal epithelial cells
Carroll DJ, Burns MWN, Mottram L, Propheter DC, Boucher A, Lessen GM, Kumar A, Malaker SA, Xing C, Hooper LV, Yrlid U, Kohler JJ. Interleukin-22 regulates B3GNT7 expression to induce fucosylation of glycoproteins in intestinal epithelial cells. Journal Of Biological Chemistry 2021, 298: 101463. PMID: 34864058, PMCID: PMC8808068, DOI: 10.1016/j.jbc.2021.101463.Peer-Reviewed Original ResearchTumor Infiltrating Lymphocytes Target HLA-I Phosphopeptides Derived From Cancer Signaling in Colorectal Cancer
Penny SA, Abelin JG, Malaker SA, Myers PT, Saeed AZ, Steadman LG, Bai DL, Ward ST, Shabanowitz J, Hunt DF, Cobbold M. Tumor Infiltrating Lymphocytes Target HLA-I Phosphopeptides Derived From Cancer Signaling in Colorectal Cancer. Frontiers In Immunology 2021, 12: 723566. PMID: 34504498, PMCID: PMC8421858, DOI: 10.3389/fimmu.2021.723566.Peer-Reviewed Original ResearchConceptsT cell responsesColorectal cancerCRC patientsPatient tumorsTumor antigensCell responsesPeripheral T cell responsesCytotoxic T-cell infiltrationFuture immunotherapeutic strategiesCRC liver metastasesNovel immunotherapeutic targetT cell infiltrationTumor-Infiltrating LymphocytesTumor-specific antigensPrimary CRC tumorsCRC cell linesKey prognostic indicatorMarker of malignancyCRC patient tumorsMultifunctional CD8Immunotherapeutic strategiesLiver metastasesInfiltrating lymphocytesCytokine responsesSame HLAMucinomics 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 ResearchSmall RNAs are modified with N-glycans and displayed on the surface of living cells
Flynn RA, Pedram K, Malaker SA, Batista PJ, Smith BAH, Johnson AG, George BM, Majzoub K, Villalta PW, Carette JE, Bertozzi CR. Small RNAs are modified with N-glycans and displayed on the surface of living cells. Cell 2021, 184: 3109-3124.e22. PMID: 34004145, PMCID: PMC9097497, DOI: 10.1016/j.cell.2021.04.023.Peer-Reviewed Original ResearchConceptsLiving cellsGlycan biosynthetic machineryDomains of lifeMultiple cell typesRNA biologySmall RNAsExtracellular biologyBiosynthetic machineryBiochemical approachesMammalian speciesBattery of chemicalAnti-dsRNA antibodiesN-glycansCell typesReceptor familyCultured cellsCell surfaceRNAThird scaffoldGlycoRNABiologyMajor targetGlycosylationGlycansSialic acidClassification, 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 microbesVectorMOD: Method for Bottom-Up Proteomic Characterization of rAAV Capsid Post-Translational Modifications and Vector Impurities
Rumachik NG, Malaker SA, Paulk NK. VectorMOD: Method for Bottom-Up Proteomic Characterization of rAAV Capsid Post-Translational Modifications and Vector Impurities. Frontiers In Immunology 2021, 12: 657795. PMID: 33868302, PMCID: PMC8047074, DOI: 10.3389/fimmu.2021.657795.Peer-Reviewed Original ResearchGenome-wide CRISPR screens reveal a specific ligand for the glycan-binding immune checkpoint receptor Siglec-7
Wisnovsky S, Möckl L, Malaker SA, Pedram K, Hess GT, Riley NM, Gray MA, Smith BAH, Bassik MC, Moerner WE, Bertozzi CR. Genome-wide CRISPR screens reveal a specific ligand for the glycan-binding immune checkpoint receptor Siglec-7. Proceedings Of The National Academy Of Sciences Of The United States Of America 2021, 118: e2015024118. PMID: 33495350, PMCID: PMC7865165, DOI: 10.1073/pnas.2015024118.Peer-Reviewed Original Research
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 experimentsCellsTargeted glycan degradation potentiates the anticancer immune response in vivo
Gray MA, Stanczak MA, Mantuano NR, Xiao H, Pijnenborg JFA, Malaker SA, Miller CL, Weidenbacher PA, Tanzo JT, Ahn G, Woods EC, Läubli H, Bertozzi CR. Targeted glycan degradation potentiates the anticancer immune response in vivo. Nature Chemical Biology 2020, 16: 1376-1384. PMID: 32807964, PMCID: PMC7727925, DOI: 10.1038/s41589-020-0622-x.Peer-Reviewed Original ResearchMeSH KeywordsAllograftsAnimalsAntibodies, MonoclonalB7-H1 AntigenCell Line, TumorHumansHydrolysisImmunoconjugatesImmunotherapyKiller Cells, NaturalMelanoma, ExperimentalMiceMice, Inbred C57BLMice, KnockoutModels, MolecularMolecular Targeted TherapyNeuraminidasePolysaccharidesProgrammed Cell Death 1 ReceptorProtein BindingProtein Interaction Domains and MotifsProtein Structure, SecondaryReceptor, ErbB-2Sialic Acid Binding Immunoglobulin-like LectinsSurvival AnalysisT-LymphocytesConceptsImmune checkpoint inhibitor therapyTumor-infiltrating myeloid cellsCheckpoint inhibitor therapyImmune cell infiltrationPowerful treatment optionAnticancer immune responseSurvival of miceSyngeneic breast cancer modelImmune cell activationBreast cancer modelBreast cancer cellsCheckpoint therapyMost patientsInhibitor therapyPD-1Checkpoint receptorsImmune suppressionTreatment optionsCell infiltrationImmune responseMyeloid cellsCancer modelCell activationCertain cancersCancer typesAn 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