RT Journal Article
SR Electronic
T1 The mechanisms of catalysis and ligand binding for the SARS-CoV-2 NSP3 macrodomain from neutron and X-ray diffraction at room temperature
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.02.07.479477
DO 10.1101/2022.02.07.479477
A1 Galen J. Correy
A1 Daniel W. Kneller
A1 Gwyndalyn Phillips
A1 Swati Pant
A1 Silvia Russi
A1 Aina E. Cohen
A1 George Meigs
A1 James M. Holton
A1 Stefan Gahbauer
A1 Michael C. Thompson
A1 Alan Ashworth
A1 Leighton Coates
A1 Andrey Kovalevsky
A1 Flora Meilleur
A1 James S. Fraser
YR 2022
UL http://biorxiv.org/content/early/2022/02/09/2022.02.07.479477.abstract
AB The NSP3 macrodomain of SARS CoV 2 (Mac1) removes ADP-ribosylation post-translational modifications, playing a key role in the immune evasion capabilities of the virus responsible for the COVID-19 pandemic. Here, we determined neutron and X-ray crystal structures of the SARS-CoV-2 NSP3 macrodomain using multiple crystal forms, temperatures, and pHs, across the apo and ADP-ribose-bound states. We characterize extensive solvation in the Mac1 active site, and visualize how water networks reorganize upon binding of ADP-ribose and non-native ligands, inspiring strategies for displacing waters to increase potency of Mac1 inhibitors. Determining the precise orientations of active site water molecules and the protonation states of key catalytic site residues by neutron crystallography suggests a catalytic mechanism for coronavirus macrodomains distinct from the substrate-assisted mechanism proposed for human MacroD2. These data provoke a re-evaluation of macrodomain catalytic mechanisms and will guide the optimization of Mac1 inhibitors.Competing Interest StatementA. Ashworth is a co-founder of Tango Therapeutics, Azkarra Therapeutics, Ovibio Corporation; a consultant for SPARC, Bluestar, ProLynx, Earli, Cura, GenVivo and GSK; a member of the SAB of Genentech, GLAdiator, Circle and Cambridge Science Corporation; receives grant/research support from SPARC and AstraZeneca; holds patents on the use of PARP inhibitors held jointly with AstraZeneca which he has benefitted financially (and may do so in the future). J. Fraser is a consultant for, has equity in, and receives research support from Relay Therapeutics.