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dc.contributor.authorTrastoy, Beatriz
dc.contributor.authorDu, Jonathan J
dc.contributor.authorLi, Chao
dc.contributor.authorGarcía-Alija, Mikel
dc.contributor.authorKlontz, Erik H
dc.contributor.authorRoberts, Blaine R
dc.contributor.authorDonahue, Thomas C
dc.contributor.authorWang, Lai-Xi
dc.contributor.authorSundberg, Eric J
dc.contributor.authorGuerin, Marcelo E
dc.date.accessioned2021-08-03T13:43:33Z
dc.date.available2021-08-03T13:43:33Z
dc.date.issued2021-07-26
dc.identifier.urihttp://hdl.handle.net/10713/16288
dc.description.abstractN-glycosylation is one of the most abundant post-translational modifications of proteins, essential for many physiological processes, including protein folding, protein stability, oligomerization and aggregation, and molecular recognition events. Defects in the N-glycosylation pathway cause diseases that are classified as congenital disorders of glycosylation. The ability to manipulate protein N-glycosylation is critical not only to our fundamental understanding of biology, but also for the development of new drugs for a wide range of human diseases. Chemoenzymatic synthesis using engineered endo-β-N-acetylglucosaminidases (ENGases) has been used extensively to modulate the chemistry of N-glycosylated proteins. However, defining the molecular mechanisms by which ENGases specifically recognize and process N-glycans remains a major challenge. Here we present the X-ray crystal structure of the ENGase EndoBT-3987 from Bacteroides thetaiotaomicron in complex with a hybrid type (Hy-type) glycan product. In combination with alanine scanning mutagenesis, molecular docking calculations and enzymatic activity measurements conducted on a chemically engineered monoclonal antibody substrate unveil two mechanisms for Hy-type recognition and processing by paradigmatic ENGases. Altogether, the experimental data provide pivotal insight into the molecular mechanism of substrate recognition and specificity for GH18 ENGases and further advance our understanding of chemoenzymatic synthesis and remodeling of homogeneous N-glycan glycoproteins.en_US
dc.description.urihttps://doi.org/10.1016/j.jbc.2021.101011en_US
dc.language.isoenen_US
dc.publisherElsevier Ltd.en_US
dc.relation.ispartofJournal of Biological Chemistryen_US
dc.rightsCopyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.en_US
dc.subjectantibody glycoengineeringen_US
dc.subjectcarbohydrate active enzymesen_US
dc.subjectendo-β-N-acetylglucosaminidasesen_US
dc.subjectenzyme specificityen_US
dc.subjectglycoprotein bioengineeringen_US
dc.subjectglycoside hydrolasesen_US
dc.subjectgut microbiomeen_US
dc.titleGH18 ENDO-β-N-ACETYLGLUCOSAMINIDASES USE DISTINCT MECHANISMS TO PROCESS HYBRID-TYPE N-LINKED GLYCANSen_US
dc.typeArticleen_US
dc.identifier.doi10.1016/j.jbc.2021.101011
dc.identifier.pmid34324829
dc.source.beginpage101011
dc.source.endpage
dc.source.countryUnited States


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