Structural basis for the recognition of complex-Type N-glycans by Endoglycosidase S

• Type of Material: Article
• Authors: Beatriz Trastoy, CIC BioGUNE; Erik Klontz, University of Maryland; Jared Orwenyo, University of Maryland; Alberto Marina, CIC BioGUNE; Lai Xi Wang, University of Maryland; Eric Sundberg, Emory University; Marcelo E. Guerin, CIC BioGUNE
• Language: English
• Date: 2018-12-01
• Publisher: Nature Research (part of Springer Nature): Fully open access journals
• Publication Version: Final Published Version
• Copyright Statement: © 2018 The Author(s).
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1038/s41467-018-04300-x
• Title of Journal or Parent Work: Nature Communications
• ISSN: 2041-1723
• Volume: 9
• Issue: 1
• Start Page: 1874
• End Page: 1874
• Grant/Funding Information: This work was supported by the MINECO Contract BFU2016-77427-C2-2-R and Severo Ochoa Excellence Accreditation (SEV-2016-0644) (to M.E.G.), Juan de la Cierva Program IJCI-2014-19206 (B.T.).
• Keywords: Oligosaccharides; Carbohydrate Sequence; Molecular Docking Simulation; Genetic Vectors; Antibodies, Monoclonal; Protein Binding; Streptomyces; Bacterial Proteins; Gene Expression; Hydrolysis; Isoenzymes; Protein Interaction Domains and Motifs; Protein Conformation, beta-Strand; Vibrio cholerae; Trichoderma; Amino Acid Motifs; Escherichia coli; Cloning, Molecular; Protein Conformation, alpha-Helical; Kinetics; Catalytic Domain; Thermodynamics; Substrate Specificity; Streptococcus pyogenes; Recombinant Proteins; Glycoside Hydrolases; Crystallography, X-Ray
• Research Categories: Health Sciences, Immunology; Chemistry, Biochemistry

Abstract

Endoglycosidase S (EndoS) is a bacterial endo-β-N-Acetylglucosaminidase that specifically catalyzes the hydrolysis of the β-1,4 linkage between the first two N-Acetylglucosamine residues of the biantennary complex-Type N-linked glycans of IgG Fc regions. It is used for the chemoenzymatic synthesis of homogeneously glycosylated antibodies with improved therapeutic properties, but the molecular basis for its substrate specificity is unknown. Here, we report the crystal structure of the full-length EndoS in complex with its oligosaccharide G2 product. The glycoside hydrolase domain contains two well-defined asymmetric grooves that accommodate the complex-Type N-linked glycan antennae near the active site. Several loops shape the glycan binding site, thereby governing the strict substrate specificity of EndoS. Comparing the arrangement of these loops within EndoS and related endoglycosidases, reveals distinct-binding site architectures that correlate with the respective glycan specificities, providing a basis for the bioengineering of endoglycosidases to tailor the chemoenzymatic synthesis of monoclonal antibodies.