Identification and characterization of AckA-dependent protein acetylation in Neisseria gonorrhoeae

• Published in: PloS one 2017-06, Vol.12 (6), p.e0179621-e0179621
• Main Authors: Post, Deborah M B, Schilling, Birgit, Reinders, Lorri M, D'Souza, Alexandria K, Ketterer, Margaret R, Kiel, Steven J, Chande, Aroon T, Apicella, Michael A, Gibson, Bradford W
• Format: Article
• Language: English
• Subjects: Acetate kinase; Acetate Kinase - genetics; Acetate Kinase - metabolism; Acetylation; Aerobic conditions; Affinity; Aging; Anaerobic conditions; Antibiotics; Assembly; Bacteria; Bacterial proteins; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biofilms; Bioinformatics; Biology and Life Sciences; Drug resistance; E coli; Enrichment; Gene Expression Regulation, Bacterial; Gonorrhea; Gram-negative bacteria; Infections; Iron; Lysine; Mass Spectrometry; Maturation; Medicine and Health Sciences; Metabolic Networks and Pathways - physiology; Metabolism; Neisseria gonorrhoeae; Neisseria gonorrhoeae - metabolism; Pathogenesis; Peptides; Phosphate; Phosphates; Phosphorylation; Physical Sciences; Physiological aspects; Post-translation; Protein Processing, Post-Translational; Proteins; Public health; Quantitation; Research and Analysis Methods; Ribosomal proteins; Salmonella; Sexually transmitted diseases; Signal transduction; Spectrometry; STD; Studies; Translation; Womens health
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0179621

Neisseria gonorrhoeae, the causative agent of gonorrhea, has a number of factors known to contribute to pathogenesis; however, a full understanding of these processes and their regulation has proven to be elusive. Post-translational modifications (PTMs) of bacterial proteins are now recognized as one mechanism of protein regulation. In the present study, Western blot analyses, with an anti-acetyl-lysine antibody, indicated that a large number of gonococcal proteins are post-translationally modified. Previous work has shown that Nε-lysine acetylation can occur non-enzymatically with acetyl-phosphate (AcP) as the acetyl donor. In the current study, an acetate kinase mutant (1291ackA), which accumulates AcP, was generated in N. gonorrhoeae. Broth cultures of N. gonorrhoeae 1291wt and 1291ackA were grown, proteins extracted and digested, and peptides containing acetylated-lysines (K-acetyl) were affinity-enriched from both strains. Mass spectrometric analyses of these samples identified a total of 2686 unique acetylation sites. Label-free relative quantitation of the K-acetyl peptides derived from the ackA and wild-type (wt) strains demonstrated that 109 acetylation sites had an ackA/wt ratio>2 and p-values