Integrative Dynamic Proteomics and Acyl-Proteomics Analyses in Clostridium butyricum

Area of Science:

• Microbiology and Molecular Biology
• Post-Translational Modifications
• Metabolomics

Background:

• Clostridium butyricum is a probiotic bacterium known for producing butyrate and acetate, crucial short-chain fatty acids (SCFAs) for gut health.
• The regulatory roles of lysine butyrylation (Kbu) and lysine acetylation (Kac), influenced by SCFAs, are not well understood in C. butyricum.
• Understanding these modifications is key to elucidating C. butyricum's beneficial functions.

Purpose of the Study:

• To conduct a comprehensive dynamic multiomic analysis of protein expression, Kbu, and Kac in C. butyricum.
• To investigate the regulatory mechanisms of Kbu during the bacterium's growth cycle.
• To identify specific proteins and sites modified by Kbu and Kac.

Main Methods:

• Dynamic multiomic analysis including proteome, Kbu, and Kac profiling.
• Bioinformatic analysis to identify functional changes and affected pathways.
• Enzymatic experiments to validate Kbu modification sites on key substrates.

Main Results:

• Identification of 2622 proteins, 1887 Kbu sites, and 2188 Kac sites.
• Dynamic changes in gene expression and energy metabolism observed at proteome and post-translational modification levels throughout the growth cycle.
• Kbu modification confirmed at active sites of alanine-tRNA ligase (K794), GMP synthase (K384), and butyrate kinase (K269).

Conclusions:

• This study provides novel insights into lysine acylations, particularly Kbu, in C. butyricum.
• Dynamic Kbu regulation plays a significant role in bacterial growth, gene expression, and energy metabolism.
• Findings enhance understanding of C. butyricum's functional mechanisms as a probiotic.