NMR side-chain assignments of the Crimean-Congo haemorrhagic fever virus glycoprotein n cytosolic domain

Abstract. We assigned the side-chain resonances of the Crimean-Congo haemorrhagic fever virus (CCHFV) glycoprotein n (Gn) cytosolic (cyto) domain to complete the backbone resonances previously published by Estrada et al. (2011). The process was accelerated by three major factors: first, sample preparation using cell-free protein synthesis (CFPS) was completed in less than 2 days and allowed correct zinc finger formation by adding zinc ions to the reaction. Second, remote access to NMR platforms with standardized pulse sequences allowed data acquisition in 18 days. Third, data analysis using the online platform NMRtist allowed sequential resonance assignments to be made in a day, verified and finalized in less than a week. Our work thus provides an example of how NMR assignments, including side chains, of small and well-behaved proteins can be approached in a rapid routine, at protein concentrations of 150 μM.