An Unprecedented Octahedral Trifluoromagnesate MgF3(Wat)– Transition State Analog Reveals The Molecular Mechanism of ATP Hydrolysis by Zika Virus Helicase

28 February 2020, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

Metal fluoride complexes mimic the transferring phosphoryl group in many enzyme-catalyzed reactions. We here employ the trifluoromagnesate transition state analog (TSA) to study a Zika virus NS3h helicase, which uses energy from ATP hydrolysis to reorganize ssRNA leading to completion of virus replication. The crystal structure of this TSA complex displays two conformations for a catalytically important loop, demonstrating how ATP hydrolysis can be coupled with RNA translocation. Unexpectedly, the trifluoromagnesate core of this transition state complex is octahedral. It is identified as having an unprecedented MgF3(Wat) ligand, confirmed by 19F NMR analysis. This structure was further probed by quantum mechanical calculations of the catalytic core (200 atoms), confirming the structural data interpretation and the concerted mechanism of ATP hydrolysis by this class of helicase. The formation of this MgF3(Wat) ligand in helicase but not in other multiple MFx structures for ATPases and GTPases strongly implies they cannot possess such an additional water in their active sites.

Keywords

Zika virus helicase
transition state analog BMP
ATPase
19F NMR spectroscopy
Protein Crystallography
General base catalysis
Phosphoryl transfer mechanism

Supplementary materials

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Chemrxiv SI 26022020
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