Methods-2023 Lysine Pulse Programs
Advances in Direct Detection of Lysine Methylation and Acetylation by Nuclear Magnetic Resonance using 13C-Enriched Cofactors
Post-translational modifications (PTMs) are reversible chemical modifications that can modulate protein structure and function. Methylation and acetylation are two such PTMs with integral and well-characterized biological roles, including modulation of chromatin structure; and unknown or poorly understood roles, exemplified by the influence of these PTMs on transcription factor structure and function. The need for biological insights into the function of these PTMs motivates the development of a nondestructive and label-free method that enables pursuit of molecular mechanisms. Here, we present a protocol for implementing nuclear magnetic resonance (NMR) methods that allow for unambiguous detection of methylation and acetylation events and demonstrate their utility by observing these marks on histone H3 tail as a model system. We leverage strategic isotopic enrichment of cofactor and peptide for visualization by [1H, 13C]-HSQC and 13C direct-detect NMR measurements. Finally, we present 13C-labeling schemes that facilitate one-dimensional NMR experiments, which combine reduced measurement time relative to two-dimensional spectroscopy with robust filtering of background signals that would otherwise create spectral crowding or limit detection of low-abundance analytes.
This collection contains the eight pulse programs and parameter sets for acquisition of the NMR data described in this paper.
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| Work Title | Methods-2023 Lysine Pulse Programs |
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| License | CC BY 4.0 (Attribution) |
| Work Type | Dataset |
| Publication Date | 2023 |
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| Deposited | July 27, 2023 |