Identification of Lysine Acetylation in Mycobacterium abscessus Using LC–MS/MS after Immunoprecipitation

Abstract

Mycobacterium abscessus (MAB), which manifests in the pulmonary system, is one of the neglected causes of nontuberculous mycobacteria (NTM) infection. Treatment against MAB is difficult, characterized by its intrinsic antibiotic drug resistance. Lysine acetylation can alter the physiochemical property of proteins in living organisms. This study aimed to determine if this protein post-translational modification (PTM) exists in a clinical isolate M. abscessus GZ002. We used the antiacetyl-lysine immunoprecipitation to enrich the low-abundant PTM proteins, followed by the LC-MS/MS analysis. The lysine acetylome of M. abscessus GZ002 was determined. There were 459 lysine acetylation sites found in 289 acetylated proteins. Lysine acetylation occurred in 5.87% of the M. abscessus GZ002 proteome, and at least 25% of them were growth essential. Aerobic respiration and carbohydrate metabolic pathways of M. abscessus GZ002 were enriched with lysine acetylation. Through bioinformatics analysis, we identified four major acetyl motif logos (K(ac)Y, K(ac)F, K(ac)H, and DK(ac)). Further comparison of the reported M. tuberculosis (MTB) acetylomes and that of MAB GZ002 revealed several common features between these two species. The lysine residues of several antibiotic-resistance, virulence, and persistence-related proteins were acetylated in both MAB GZ002 and MTB. There were 51 identical acetylation sites in 37 proteins found in common between MAB GZ002 and MTB. Overall, we demonstrate a profile of lysine acetylation in MAB GZ002 proteome that shares similarities with MTB. Interventions that target at these conserved sections may be valuable as anti-NTM or anti-TB therapies.

Comparative Proteomic Analysis of Lysine Acetylation in Fish CIK Cells Infected with Aquareovirus

“…These results suggested that GCRV infection induced wide changes in host cellular proteins at lysine acetylation level. Additionally, many lysine acetylation motifs identified in this study were also found in other eukaryotic and prokaryotic species [ 37 , 38 , 39 , 40 , 41 , 42 ], indicating that the lysine acetylation motifs are highly conserved among different species.…”

Functional Insights Into Protein Acetylation in the Hyperthermophilic Archaeon Sulfolobus islandicus*

“…Lys and Arg) are enriched at the ϩ1 and ϩ2 positions, in relation to the residue of acetylation, in the acetylomes of the two species. In comparison, Tyr is significantly enriched at the ϩ1 position whereas Lys or Arg is always located downstream of the ϩ2 position in the mycobacterium, drosophila and human acetylomes (12,13,92,93). H. mediterranei encodes twelve acyl-CoA synthetases.…”

Assessment of Clofazimine and TB47 Combination Activity against Mycobacterium abscessus Using a Bioluminescent Approach

“…Escherichia coli DH5␣ was grown at 37°C in Luria-Bertani (LB) medium. M. abscessus GZ002, a clinical isolate described previously (30), for which the whole genome and DNA methylome have been sequenced and analyzed (31), was used as the parental strain to construct the UAlMab strain. In addition, 20 new clinical strains of M. abscessus subsp.…”

“…In addition, 20 new clinical strains of M. abscessus subsp. abscessus isolated from Guangzhou Chest Hospital (Guangzhou, China), as identified by comparison of sequences of the 16S rRNA gene, the DNA gyrase subunit gene, and the DNA-directed RNA polymerase subunit ␤ gene ( Table 2 gives primer information), with their corresponding homologous genes in the reference strain M. abscessus ATCC 19977 (30), were investigated in this study. M. abscessus strains were grown at 37°C in Middlebrook 7H9 broth (Difco) supplemented with 0.05% Tween 80 and 10% oleic acid-albumin-dextrose-catalase (OADC; Difco), or on Middlebrook 7H11 agar (Difco) supplemented with 10% OADC.…”