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 Sakhrani; Leighton Coates; Chad M. Rienstra; Leonard J. Mueller

doi:10.1007/s12104-025-10242-5

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 Sakhrani, Leighton Coates, Chad M. Rienstra, Leonard J. Mueller

STS Project Technical Systems

Research output: Contribution to journal › Article › peer-review

Abstract

¹³C and ¹⁵N backbone chemical shift assignments are reported for the 28.5 kDa protein Toho-1 β-lactamase, a Class A extended spectrum β-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 side-chain chemical shifts and catalytic mechanism.

Original language	English
Pages (from-to)	245-254
Number of pages	10
Journal	Biomolecular NMR Assignments
Volume	19
Issue number	2
DOIs	https://doi.org/10.1007/s12104-025-10242-5
State	Published - Dec 2025

Funding

This study made use of NMRFAM, a National Institutes of Health Biomedical Technology Development and Dissemination Center (P41GM136463). The 1.1 GHz NMR spectrometer was funded by the United States National Science Foundation Mid-Scale Research Infrastructure Big Idea (1946970). Helium recovery equipment, computers, and infrastructure for data archive were funded by the University of Wisconsin-Madison, National Institutes of Health (P41GM136463, R24GM141526), and National Science Foundation (1946970). L.J.M. was supported by grants from the National Institutes of Health (R01GM137008 and R35GM145369).

Keywords

1.1 GHz NMR
Backbone chemical shift assignments
Protein solid-state NMR
Toho-1 β-lactamase

Access to Document

10.1007/s12104-025-10242-5

Cite this

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title = "Backbone assignment of a 28.5 kDa class A extended spectrum β-lactamase by high-field, carbon-detected solid-state NMR",

abstract = "13C and 15N backbone chemical shift assignments are reported for the 28.5 kDa protein Toho-1 β-lactamase, a Class A extended spectrum β-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 side-chain chemical shifts and catalytic mechanism.",

keywords = "1.1 GHz NMR, Backbone chemical shift assignments, Protein solid-state NMR, Toho-1 β-lactamase",

author = "Williams, \{Christopher G.\} and Songlin Wang and Thome, \{Alexander F.\} and Warmuth, \{Owen A.\} and Varun Sakhrani and Leighton Coates and Rienstra, \{Chad M.\} and Mueller, \{Leonard J.\}",

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T1 - Backbone assignment of a 28.5 kDa class A extended spectrum β-lactamase by high-field, carbon-detected solid-state NMR

AU - Williams, Christopher G.

AU - Wang, Songlin

AU - Thome, Alexander F.

AU - Warmuth, Owen A.

AU - Sakhrani, Varun

AU - Coates, Leighton

AU - Rienstra, Chad M.

AU - Mueller, Leonard J.

N1 - Publisher Copyright: © The Author(s), under exclusive licence to Springer Nature B.V. 2025.

PY - 2025/12

Y1 - 2025/12

N2 - 13C and 15N backbone chemical shift assignments are reported for the 28.5 kDa protein Toho-1 β-lactamase, a Class A extended spectrum β-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 side-chain chemical shifts and catalytic mechanism.

AB - 13C and 15N backbone chemical shift assignments are reported for the 28.5 kDa protein Toho-1 β-lactamase, a Class A extended spectrum β-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 side-chain chemical shifts and catalytic mechanism.

KW - 1.1 GHz NMR

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KW - Protein solid-state NMR

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