ScholarSphere

NMR Data for Dewing and Showalter 2025

All NMR data that are represented in the acylation time courses are included in this upload. The raw data are presented as collected in the Bruker Topspin environment.

Title: Probing Enzymatic Acetylation Events in Real-Time with NMR Spectroscopy: Insights into Acyl-Cofactor Dependent p300 Modification of Histone H4

https://doi.org/10.1002/prot.26848

Abstract: Lysine acylation is a rapidly expanding class of post-translational modifications with largely unexplored functional roles; the study of acylations beyond acetylation is especially impeded by limited methods for their preparation, detection, and characterization in vitro. We previously reported a nuclear magnetic resonance (NMR)-based approach to monitor Nε-lysine acetylation following Ada2/Gcn5-catalyzed installation of a 13C-acetyl probe on the histone H3 tail. Building on this foundation, here we expand those techniques by demonstrating the installation and 1H, 13C-HSQC based NMR detection of both 13C-acetyl and 13C-propionyl probes on the histone H4 tail using a mutant p300 lysine acetyltransferase (KAT) enzyme with enhanced activity. Additionally, we introduce a continuous evaluation method for acyltransferase reaction data, enabling the extraction of relative rate constants—a technique inspired by our laboratory’s recent work on NMR methyltransferase kinetics. This study demonstrates that our NMR-based approach to assay enzymatic 13C-acylation is adaptable, providing a versatile platform for investigating a range of acylations, KAT enzymes, and protein substrates. Notably, in the process of developing these methods, we observed that p300 KAT may display distinct modification site preferences and regulatory mechanisms depending on the acyl cofactor utilized, underscoring the method’s potential to advance the emerging field of lysine acylation biochemistry.

Citation

SHOWALTER, SCOTT A; Dewing, Sophia (2025). NMR Data for Dewing and Showalter 2025 [Data set]. Scholarsphere.

Files

Metadata

Work Title NMR Data for Dewing and Showalter 2025
Subtitle p300 acylation time course data
Access
Open Access
Creators
  1. SCOTT A SHOWALTER
  2. Sophia M Dewing
Keyword
  1. p300
  2. acylation
  3. acetylation
  4. nuclear magnetic resonance
  5. kinetics
License CC BY 4.0 (Attribution)
Work Type Dataset
Acknowledgments
  1. R21-GM137129
  2. T32-GM102057
Publisher
  1. Scott Showalter
Publication Date 2025
Subject
  1. protein biophysics
Language
  1. English
Geographic Area
  1. United States
Related URLs
  1. https://doi.org/10.1002/prot.26848
Source
  1. Scott Showalter
Deposited June 10, 2025

Versions

Analytics

Collections

This resource is currently not in any collection.

Work History

Version 1
published

  • Created

    June 10, 2025 09:32 by sas76

  • Updated

    June 10, 2025 09:32 by [unknown user]

  • Updated Source, Keyword, Subject, and 7 more Show Changes

    June 10, 2025 09:40 by sas76

    Source
    • Scott Showalter
    Keyword
    • p300, acylation, acetylation, nuclear magnetic resonance, kinetics
    Subject
    • protein biophysics
    Language
    • English
    Publisher
    • Scott Showalter
    Geographic Area
    • United States
    Related URLs
    • https://doi.org/10.1002/prot.26848
    Subtitle
    • p300 acylation time course data
    Description
    • All NMR data that are represented in the acylation time courses are included in this upload. The raw data are presented as collected in the Bruker Topspin environment.
    • Title: Probing Enzymatic Acetylation Events in Real-Time with NMR Spectroscopy: Insights into Acyl-Cofactor Dependent p300 Modification of Histone H4
    • https://doi.org/10.1002/prot.26848
    • Abstract: Lysine acylation is a rapidly expanding class of post-translational modifications with largely unexplored functional roles; the study of acylations beyond acetylation is especially impeded by limited methods for their preparation, detection, and characterization in vitro. We previously reported a nuclear magnetic resonance (NMR)-based approach to monitor Nε-lysine acetylation following Ada2/Gcn5-catalyzed installation of a 13C-acetyl probe on the histone H3 tail. Building on this foundation, here we expand those techniques by demonstrating the installation and 1H, 13C-HSQC based NMR detection of both 13C-acetyl and 13C-propionyl probes on the histone H4 tail using a mutant p300 lysine acetyltransferase (KAT) enzyme with enhanced activity. Additionally, we introduce a continuous evaluation method for acyltransferase reaction data, enabling the extraction of relative rate constants—a technique inspired by our laboratory’s recent work on NMR methyltransferase kinetics. This study demonstrates that our NMR-based approach to assay enzymatic 13C-acylation is adaptable, providing a versatile platform for investigating a range of acylations, KAT enzymes, and protein substrates. Notably, in the process of developing these methods, we observed that p300 KAT may display distinct modification site preferences and regulatory mechanisms depending on the acyl cofactor utilized, underscoring the method’s potential to advance the emerging field of lysine acylation biochemistry.
    Publication Date
    • 2025
  • Updated Acknowledgments Show Changes

    June 10, 2025 09:41 by sas76

    Acknowledgments
    • R21-GM137129, T32-GM102057
  • Added Creator SCOTT A SHOWALTER

    June 10, 2025 09:41 by sas76

  • Added Creator Sophia M Dewing

    June 10, 2025 09:41 by sas76

  • Added AcTimecourses.zip

    June 10, 2025 11:32 by sas76

  • Added README.md

    June 10, 2025 11:32 by sas76

  • Updated License Show Changes

    June 10, 2025 11:32 by sas76

    License
    • https://creativecommons.org/licenses/by/4.0/
  • Published

    June 10, 2025 11:32 by sas76

  • Updated

    June 10, 2025 22:05 by [unknown user]