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dc.contributor.authorKlontz, E.H.
dc.contributor.authorLi, C.
dc.contributor.authorKihn, K.
dc.contributor.authorFields, J.K.
dc.contributor.authorBeckett, D.
dc.contributor.authorSnyder, G.A.
dc.contributor.authorWintrode, P.L.
dc.contributor.authorDeredge, D.
dc.contributor.authorWang, L.-X.
dc.contributor.authorSundberg, E.J.
dc.date.accessioned2021-02-08T20:13:24Z
dc.date.available2021-02-08T20:13:24Z
dc.date.issued2020-12-04
dc.identifier.urihttp://hdl.handle.net/10713/14543
dc.description.abstractFucosylation is important for the function of many proteins with biotechnical and medical applications. Alpha-fucosidases comprise a large enzyme family that recognizes fucosylated substrates with diverse α-linkages on these proteins. Lactobacillus casei produces an α-fucosidase, called AlfC, with specificity towards α(1,6)-fucose, the only linkage found in human N-glycan core fucosylation. AlfC and certain point mutants thereof have been used to add and remove fucose from monoclonal antibody N-glycans, with significant impacts on their effector functions. Despite the potential uses for AlfC, little is known about its mechanism. Here, we present crystal structures of AlfC, combined with mutational and kinetic analyses, hydrogen–deuterium exchange mass spectrometry, molecular dynamic simulations, and transfucosylation experiments to define the molecular mechanisms of the activities of AlfC and its transfucosidase mutants. Our results indicate that AlfC creates an aromatic subsite adjacent to the active site that specifically accommodates GlcNAc in α(1,6)-linkages, suggest that enzymatic activity is controlled by distinct open and closed conformations of an active-site loop, with certain mutations shifting the equilibrium towards open conformations to promote transfucosylation over hydrolysis, and provide a potentially generalizable framework for the rational creation of AlfC transfucosidase mutants. Copyright 2020, The Author(s).en_US
dc.description.sponsorshipThis research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02-06CH11357.en_US
dc.description.urihttps://doi.org/10.1038/s41467-020-20044-zen_US
dc.language.isoen_USen_US
dc.publisherNature Researchen_US
dc.relation.ispartofNature Communications
dc.subject.meshalpha-L-Fucosidaseen_US
dc.titleStructure and dynamics of an α-fucosidase reveal a mechanism for highly efficient IgG transfucosylationen_US
dc.typeArticleen_US
dc.identifier.doi10.1038/s41467-020-20044-z
dc.identifier.pmid33277506


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