6E58

Crystal structure of Streptococcus pyogenes endo-beta-N-acetylglucosaminidase (EndoS2)

Summary for 6E58

• Entry DOI: 10.2210/pdb6e58/pdb
• Descriptor: Secreted Endo-beta-N-acetylglucosaminidase (EndoS), CALCIUM ION (3 entities in total)
• Functional Keywords: glycoside hydrolase, endo-beta-n-acetylglucosaminidase s2, endoglycosidase s2, endos2, hydrolase
• Biological source: Streptococcus pyogenes M49 591
• Total number of polymer chains: 2
• Total formula weight: 181844.70

Authors

Klontz, E.H.,Trastoy, B.,Gunther, S.,Guerin, M.E.,Sundberg, E.J. (deposition date: 2018-07-19, release date: 2019-02-06, Last modification date: 2023-10-11)

Primary citation

Klontz, E.H.,Trastoy, B.,Deredge, D.,Fields, J.K.,Li, C.,Orwenyo, J.,Marina, A.,Beadenkopf, R.,Gunther, S.,Flores, J.,Wintrode, P.L.,Wang, L.X.,Guerin, M.E.,Sundberg, E.J.
Molecular Basis of Broad SpectrumN-Glycan Specificity and Processing of Therapeutic IgG Monoclonal Antibodies by Endoglycosidase S2.
ACS Cent Sci, 5:524-538, 2019

PubMed Abstract: Immunoglobulin G (IgG) glycosylation critically modulates antibody effector functions. secretes a unique endo-β--acetylglucosaminidase, EndoS2, which deglycosylates the conserved -linked glycan at Asn297 on IgG Fc to eliminate its effector functions and evade the immune system. EndoS2 and specific point mutants have been used to chemoenzymatically synthesize antibodies with customizable glycosylation for gain of functions. EndoS2 is useful in these schemes because it accommodates a broad range of -glycans, including high-mannose, complex, and hybrid types; however, its mechanism of substrate recognition is poorly understood. We present crystal structures of EndoS2 alone and bound to complex and high-mannose glycans; the broad -glycan specificity is governed by critical loops that shape the binding site of EndoS2. Furthermore, hydrolytic experiments, domain-swap chimeras, and hydrogen-deuterium exchange mass spectrometry reveal the importance of the carbohydrate-binding module in the mechanism of IgG recognition by EndoS2, providing insights into engineering enzymes to catalyze customizable glycosylation reactions.

PubMed: 30937380
DOI: 10.1021/acscentsci.8b00917

Experimental method

X-RAY DIFFRACTION (2.75 Å)