Conformational Control of Fast Asparagine Deamidation in a Norovirus Capsid Protein

Accelerated spontaneous deamidation of asparagine 373 and subsequent conversion into an isoaspartate has been shown to attenuate the binding of histo blood group antigens (HBGAs) to the protruding domain (P-domain) of the capsid protein of a prevalent norovirus strain (GII.4). Here, we link an unusual backbone conformation of asparagine 373 to its fast site-specific deamidation. NMR spectroscopy and ion exchange chromatography have been used to monitor the deamidation reaction of P-domains of two closely related GII.4 norovirus strains, specific point mutants, and control peptides. MD simulations over several microseconds have been instrumental to rationalize the experimental findings. While conventional descriptors such as available surface area, root mean-square fluctuations, or nucleophilic attack distance fail as explanations, the population of a rare syn-backbone conformation distinguishes asparagine 373 from all other asparagine residues. We suggest that stabilization of this unusual conformation enhances the nucleophilicity of the backbone nitrogen of aspartate 374, in turn accelerating the deamidation of asparagine 373. This finding should be relevant to the development of reliable prediction algorithms for sites of rapid asparagine deamidation in proteins.

Automated summary

The accelerated deamidation of asparagine 373 into isoaspartate is found to weaken the binding of histo blood group antigens (HBGAs) to the capsid protein of a prevalent norovirus strain. Unusual backbone conformation of asparagine 373 is linked to its fast site-specific deamidation. Molecular dynamics simulations and experimental techniques have shed light on this phenomenon, suggesting that stabilization of the rare syn-backbone conformation enhances the nucleophilicity of the neighboring aspartate residue and accelerates the deamidation process.