Assessment of a large enzyme‐drug complex by proton‐detected solid‐state NMR without deuteration

Solid‐state NMR has recently enabled structural biology based on small amounts of non‐deuterated proteins, largely relieving the classical sample production demands. Still, despite the benefits for sample preparation, successful and comprehensive characterization of complex spin systems in the few cases of higher molecular‐weight proteins have so far relied on traditional 13C‐detected methodology or sample deuteration. With a 29 kDa carbonic anhydrase:acetazolamide complex, we show that different aspects of solid‐state NMR assessment of a complex spin system can be successfully performed using a non‐deuterated, 500 µg sample in combination with adequate spectroscopic tools. The access to protein structure, protein dynamics, as well as biochemical parameters in amino acid sidechains like histidine protonation states, will be transferable to proteins that are not expressible in E. coli.

from A via a.sfakia on Inoreader http://bit.ly/2sPOrjP