Characterization of Critical Determinants of ACE2–SARS CoV-2 RBD Interaction

• Main Authors: Emily E. F. Brown, Reza Rezaei, Taylor R. Jamieson, Jaahnavi Dave, Nikolas T. Martin, Ragunath Singaravelu, Mathieu J. F. Crupi, Stephen Boulton, Sarah Tucker, Jessie Duong, Joanna Poutou, Adrian Pelin, Hamed Yasavoli-Sharahi, Zaid Taha, Rozanne Arulanandam, Abera Surendran, Mina Ghahremani, Bradley Austin, Chantal Matar, Jean-Simon Diallo, John C. Bell, Carolina S. Ilkow, Taha Azad
• Format: Article
• Language: English
• Published: MDPI AG 2021-02-01
• Series: International Journal of Molecular Sciences
• Subjects: SARS-CoV-2; angiotensin-converting enzyme 2; receptor binding domain; NanoLuc Binary Technology; spike protein; vaccine development
• Online Access: https://www.mdpi.com/1422-0067/22/5/2268

Despite sequence similarity to SARS-CoV-1, SARS-CoV-2 has demonstrated greater widespread virulence and unique challenges to researchers aiming to study its pathogenicity in humans. The interaction of the viral receptor binding domain (RBD) with its main host cell receptor, angiotensin-converting enzyme 2 (ACE2), has emerged as a critical focal point for the development of anti-viral therapeutics and vaccines. In this study, we selectively identify and characterize the impact of mutating certain amino acid residues in the RBD of SARS-CoV-2 and in ACE2, by utilizing our recently developed NanoBiT technology-based biosensor as well as pseudotyped-virus infectivity assays. Specifically, we examine the mutational effects on RBD-ACE2 binding ability, efficacy of competitive inhibitors, as well as neutralizing antibody activity. We also look at the implications the mutations may have on virus transmissibility, host susceptibility, and the virus transmission path to humans. These critical determinants of virus–host interactions may provide more effective targets for ongoing vaccines, drug development, and potentially pave the way for determining the genetic variation underlying disease severity.