2023
Bump-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
2020
Bump-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 ResearchMeSH KeywordsBiosynthetic PathwaysCell MembraneGlycosylationGlycosyltransferasesHEK293 CellsHep G2 CellsHumansK562 CellsN-AcetylgalactosaminyltransferasesPolysaccharidesProtein EngineeringProteinsConceptsLiving 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
2019
Engineering Orthogonal Polypeptide GalNAc-Transferase and UDP-Sugar Pairs
Choi J, Wagner LJS, Timmermans SBPE, Malaker SA, Schumann B, Gray MA, Debets MF, Takashima M, Gehring J, Bertozzi CR. Engineering Orthogonal Polypeptide GalNAc-Transferase and UDP-Sugar Pairs. Journal Of The American Chemical Society 2019, 141: 13442-13453. PMID: 31373799, PMCID: PMC6813768, DOI: 10.1021/jacs.9b04695.Peer-Reviewed Original ResearchMeSH KeywordsAcetylgalactosamineAmino Acid SequenceCatalytic DomainHumansIsoenzymesKineticsModels, MolecularMutagenesisN-AcetylgalactosaminyltransferasesProtein EngineeringSubstrate SpecificityUridine Diphosphate