Backbone Assignment of a 28.5 kDa Class A Extended Spectrum β-Lactamase by High-Field, Carbon-Detected Solid-State NMR

Christopher G. Williams; Songlin Wang; Alexander F. Thome; Owen A. Warmuth; Varun V. Sakhrani; Leighton Coates; Chad M. Rienstra; Leonard J. Mueller

doi:10.1007/s12104-025-10242-5

BiochemistryBiomolecular NMR AssignmentsJul 27, 2025

Peer-reviewed journal article

Backbone Assignment of a 28.5 kDa Class A Extended Spectrum β-Lactamase by High-Field, Carbon-Detected Solid-State NMR

Christopher G. Williams iD, Songlin Wang iD, Alexander F. Thome, Owen A. Warmuth iD, Varun V. Sakhrani, Leighton Coates iD, Chad M. Rienstra iD, Leonard J. Mueller iD

Abstract

13C and 15N backbone chemical shift assignments are reported for the 28.5 kDa protein Toho-1 {beta}-lactamase, a Class A extended spectrum {beta}-lactamase. A very high level of assignment completeness (97% of the backbone) is enabled by the combined sensitivity and resolution gains of ultrahigh-field NMR spectroscopy (1.1 GHz), improved probe technology, and optimized pulse sequences. The assigned chemical shifts agree well with our previous solution-state NMR assignments, indicating that the secondary structure is conserved in the solid state. These assignments provide a foundation for future investigations of sidechain chemical shifts and catalytic mechanism.

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