Yang Y, Du Y, Kaltashov IA

The emergence and rapid proliferation of the novel coronavirus (SARS-CoV-2) resulted in a global pandemic, with over six million cases and nearly four hundred thousand deaths reported worldwide by the end of May 2020. A rush to find the cure prompted re-evaluation of a range of existing therapeutics vis-a-vis their ...

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potential role in treating COVID-19, placing a pre-mium on analytical tools capable of supporting such efforts. Native mass spectrometry (MS) has long been a tool of choice in supporting the mechanistic studies of drug/therapeutic target interactions, but its applications remain limited in the cases that involve systems with a high level of structural heterogeneity. Both SARS-CoV-2 spike protein (S-protein), a critical ele-ment of the viral entry to the host cell, and ACE2, its docking site on the host cell surface, are extensively glycosylated, mak-ing them challenging targets for native MS. However, supplementing native MS with a gas-phase ion manipulation tech-nique (limited charge reduction) allows meaningful information to be obtained on the non-covalent complexes formed by ACE2 and the receptor-binding domain (RBD) of the S-protein. Using this technique in combination with molecular model-ing also allows the role of heparin in destabilizing the ACE2/RBD association to be studied, providing critical information for understanding the molecular mechanism of its interference with the virus docking to the host cell receptor. Both short (pen-tasaccharide) and relatively long (eicosasaccharide) heparin oligomers form 1:1 complexes with RBD, indicating the pres-ence of a single binding site. This association alters the protein conformation (to maximize the contiguous patch of the posi-tive charge on the RBD surface), resulting in a notable decrease of its ability to associate with ACE2. The destabilizing effect of heparin is more pronounced in the case of the longer chains due to the electrostatic repulsion between the low-pI ACE2 and the heparin segments not accommodated on the RBD surface. In addition to providing important mechanistic infor-mation on attenuation of the ACE2/RBD association by heparin, the study demonstrates the yet untapped potential of na-tive MS coupled to gas-phase ion chemistry as a means of facilitating rational repurposing of the existing medicines for treating COVID-19.

Date: Jul. 17, 2020

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