Antimicrobial Peptide BCp12 Inhibits Staphylococcus aureus Growth by Altering Lysine Malonylation Levels in the Arginine Synthesis Pathway

Abstract

To adapt to external stimuli, bacteria fine-tune important protein activities using post-translational modifications. The present study provides novel insights into the molecular mechanism of the antimicrobial peptide BCp12. We demonstrate that BCp12 significantly suppressed bacterial growth, induced cell apoptosis, and modulated overall malonylation levels in Staphylococcus aureus cells. Malonylateomic analysis was performed to identify the proteins malonylated by the BCp12 treatment of S. aureus. In total, 53 malonylated proteins (17 up-regulated, 36 down-regulated) were identified as differentially expressed malonylated proteins (DMPs; > 1.5-fold or <0.67-fold, P < 0.05). This result was confirmed via the identification of 21 differential metabolites (DMs; VIP > 1, P < 0.05) in the arginine and proline metabolome. Bioinformatic analysis revealed that the DMPs and DMs were especially enriched in the arginine synthesis pathway. By integrating our lysine malonylational and metabolomic data, we provide new insights into the mechanism by which BCp12 inhibits S. aureus.

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“…Peptides were separated with a gradient from 4% to 22% in solvent B (0.1% formic acid in acetonitrile) over 92 min, 22 to 30% in 20 min, and climbing to 80% in 4 min, and then held at 80% for the last 4 min (the routine analysis is usually 60 min). All steps were performed at a constant flow rate of 450 nL/min on a nanoElute UHPLC system (Bruker Daltonics, Germany) . The peptides were subjected to a capillary ion source, followed by timsTOF Pro 2 MS (Bruker Daltonics).…”

Insight into Ionic Strength-Induced Solubilization of Myofibrillar Proteins from Silver Carp (Hypophthalmichthys molitrix): Structural Changes and 4D Label-Free Proteomics Analysis

“…Subsequently, samples were digested overnight with trypsin at a 1:50 (w/w) ratio. Dithiothreitol was added at a final concentration of 5 mM for 30 min at 56 °C and alkylated with 11 mM iodoacetamide for 15 min at room temperature in darkness . Finally, the peptides were desalted and dried by vacuum freezing.…”

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