A Comprehensive Analysis of the Lysine Acetylome in the Aquatic Animals Pathogenic Bacterium Vibrio mimicus

Abstract

Protein lysine acetylation is an evolutionarily conserved post-translational modification (PTM), which is dynamic and reversible, playing a crucial regulatory role in almost every aspect of metabolism, of both eukaryotes and prokaryotes. Several global lysine acetylome studies have been carried out in various bacteria, but thus far, there have been no reports of lysine acetylation for the commercially important aquatic animal pathogen Vibrio mimicus. In the present study, we used anti-Ac-K antibody beads to highly sensitive immune-affinity purification and combined high-resolution LC-MS/MS to perform the first global lysine acetylome analysis in V. mimicus, leading to the identification of 1,097 lysine-acetylated sites on 582 proteins, and more than half (58.4%) of the acetylated proteins had only one site. The analysis of acetylated modified peptide motifs revealed six significantly enriched motifs, namely, KacL, KacR, L(-2) KacL, LKacK, L(-7) EKac, and IEKac. In addition, bioinformatic assessments state clearly that acetylated proteins have a hand in many important biological processes in V. mimicus, such as purine metabolism, ribosome, pyruvate metabolism, glycolysis/gluconeogenesis, the TCA cycle, and so on. Moreover, 13 acetylated proteins were related to the virulence of V. mimicus. To sum up, this is a comprehensive analysis whole situation protein lysine acetylome in V. mimicus and provides an important foundation for in-depth study of the biological function of lysine acetylation in V. mimicus.

Acetylome and Succinylome Profiling of Edwardsiella tarda Reveals Key Roles of Both Lysine Acylations in Bacterial Antibiotic Resistance

“…Similarly, in the cellular component (CC) category, most of the identified lysine-acetylated and -succinylated proteins were prevalent in the cell, cell part, cytoplasm, intracellular part, and intracellular categories ( Figure 3 C). Consistent with previous research, the identified Kac proteins were enriched in metabolic processes and cell locations similar to BW25113 and V. mimicus [ 29 , 48 ]. When compared to Kac proteins, the significantly enriched categories were quite different for Ksu proteins in the molecular function (MF) category.…”

“…The identified Ksu (692 proteins) and Kac proteins (588 proteins) accounted for 19.3% and 16.4% of the total proteins (3580) in E. tarda , respectively. When compared to the bacterial acetylome and succinylome of other animal pathogens studied, the percentage of Kac proteins was lower than those of V. alginolyticus (27.1%) [ 14 ] and A. hydrophila (24.6%) [ 16 ] but was higher than in V. parahemolyticus (13.6%) and V. mimicus (15.5%) [ 29 ]. The percentage of Ksu proteins was lower than in E. coli (23.91%) [ 30 ] but was higher than in several species of bacteria such as A. hydrophila (16.0%) [ 28 ], V. parahemolyticus (13.3%) [ 15 ], M. tuberculosis (17.07%) [ 31 ], and Pseudomonas aeruginosa (10.5%) [ 32 ].…”

“…Motif KacH has been reported to be conserved and is widespread in other bacterial species such as Streptococcus mutans [ 38 ], S. pneumoniae [ 39 ], Mycobacterium abscessus [ 40 ], Saccharopolyspora erythraea [ 41 ], and M. tuberculosis [ 42 ]. Motif KsuL has been reported to be enriched in B. subtilis [ 43 , 44 ], and Motif KacL has been reported in V. mimicus [ 29 ]. However, the other motifs, Ksu H, EKsu L, EKac L, V.Ksu ….K, and D…..R.Ksu, have rarely been identified in other bacteria.…”

Holistic analysis of lysine acetylation in aquaculture pathogenic bacteria Vibrio alginolyticus under bile salt stress

“…The whole protein (500 μg) of V. alginolyticus under bile salt stress was incubated with specific polyclonal antibodies OmpN, OmpR, and GrpE at 4°C overnight to precipitate target proteins, as previously described ( 10 ). Briefly, the protein A/G beads were washed three times with pre-cooled PBS buffer (137 mM NaCl, 2.7 mM KCl, 4.3 mM Na 2 HPO 4 , 1.4 mM KH 2 PO 4 , pH 7.2–7.4) and then added to the lysate for incubation at 4°C for 2 h. The beads were centrifuged at 4°C and 2,500 g for 5 min to remove the supernatant, and washed five times with pre-cooled PBS buffer.…”

Functional proteomics identify mannitol metabolism in serum resistance and therapeutic implications in Vibrio alginolyticus

“…g ., glucose, re-sensitizes antibiotic-resistant bacteria to antibiotics ( 25 , 43 ). Recent studies also show that the glycolytic pathway and the pyruvate cycle are subjected to the regulation of acetylation and succinylation, indicating that protein post-translation modification may be involved in serum resistance ( 44 – 46 ). Actually, our previous study demonstrated that the activity of the pyruvate cycle was decreased by serum ( 17 ).…”