Structure-guided glyco-engineering of ACE2 for improved potency as soluble SARS-CoV-2 decoy receptor
Tümay Capraz, Nikolaus F. Kienzl, Elisabeth Laurent, Jan Walther Perthold, Esther Föderl-Höbenreich, Clemens Grünwald‐Gruber, Daniel Maresch, Vanessa Monteil, Janine Niederhöfer, Gerald Wirnsberger, Alì Mirazimi, Kurt Zatloukal, Lukas Mach, Josef Penninger, Chris Oostenbrink, Johannes Stadlmann
Abstract
Infection and viral entry of SARS-CoV-2 crucially depends on the binding of its Spike protein to angiotensin converting enzyme 2 (ACE2) presented on host cells. Glycosylation of both proteins is critical for this interaction. Recombinant soluble human ACE2 can neutralize SARS-CoV-2 and is currently undergoing clinical tests for the treatment of COVID-19. We used 3D structural models and molecular dynamics simulations to define the ACE2 N-glycans that critically influence Spike-ACE2 complex formation. Engineering of ACE2 N-glycosylation by site-directed mutagenesis or glycosidase treatment resulted in enhanced binding affinities and improved virus neutralization without notable deleterious effects on the structural stability and catalytic activity of the protein. Importantly, simultaneous removal of all accessible N-glycans from recombinant soluble human ACE2 yields a superior SARS-CoV-2 decoy receptor with promise as effective treatment for COVID-19 patients.