Expanding the Toolset of Biomolecular NMR with Efficient and Cost-Effective ¹⁷O-Labeling via Bacterial Expression.

ABSTRACT: Oxygen plays a central role in biomolecular structures and functions, with ¹⁷O NMR emerging as a powerful tool for elucidating biomolecular properties. However, the low natural abundance of the NMR-active isotope, ¹⁷O (0.0373 %), presents a significant hurdle to its widespread application. Here, we introduce a rapid and cost-effective approach for amino acid-specific ¹⁷O-labeling of recombinant proteins. Using a common bacterial expression system and with a 30-minute rapid synthesis protocol of ¹⁷O-labeled amino acids via mechanochemical saponification, we have generated Leu- and Phe-specific ¹⁷O-labeled recombinant proteins derived from diverse organisms, including CrgA and FtsQ from Mycobacterium tuberculosis and E protein from SARS-

SUBMITTER: Tian Z 

PROVIDER: S-EPMC11724747 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature