Related Experiment Video
Updated: May 2, 2026
A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli
Published on: December 9, 2017
Protocol for identifying OXCT1-mediated LACTB succinylation sites in vitro
Wenhao Ma1, Yuchen Sun1, Huafeng Zhang2
1Anhui Key Laboratory of Hepatopancreatobiliary Surgery, Department of General Surgery, Anhui Provincial Hospital, The First Affiliated Hospital of USTC, Division of Life Science and Medicine, University of Science and Technology of China, Hefei, Anhui 230027, China; The Chinese Academy of Sciences Key Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Science and Medicine, University of Science and Technology of China, Hefei, Anhui 230027, China; Anhui Province Key Laboratory of Biomedical Aging Research, Division of Life Science and Medicine, University of Science and Technology of China, Hefei, Anhui 230027, China.
This study introduces a new protocol to measure succinylation, a key protein modification. The method uses western blotting and mass spectrometry to detect succinylation levels and sites on LACTB protein, mediated by OXCT1.
Area of Science:
- Biochemistry
- Proteomics
- Molecular Biology
Background:
- Protein succinylation is a crucial post-translational modification.
- Oxidative 1-3-hydroxybutyrate carrier protein 1 (OXCT1) functions as a succinyltransferase.
- Serine beta-lactamase-like protein (LACTB) is a target of OXCT1-mediated succinylation.
Purpose of the Study:
- To present a detailed protocol for quantifying OXCT1-mediated LACTB succinylation.
- To identify specific succinylation sites on LACTB.
- To establish a method applicable to other proteins.
Main Methods:
- Western blotting (WB) for detecting succinylation levels.
- Mass spectrometry (MS) for identifying succinylation sites.
- In vitro assays to study OXCT1-LACTB interactions.
Main Results:
- Successful detection of OXCT1-mediated LACTB succinylation levels.
- Identification of K284 as a succinylation site on LACTB.
- Validation of the protocol's applicability to other proteins.
Conclusions:
- The developed protocol enables precise measurement of protein succinylation.
- This method aids in understanding the role of OXCT1 in LACTB modification.
- The protocol serves as a valuable tool for proteomic research.
More Related Videos
12:49Quantification of Site-specific Protein Lysine Acetylation and Succinylation Stoichiometry Using Data-independent Acquisition Mass Spectrometry
Published on: April 4, 2018
12:11Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization
Published on: February 27, 2020
