Proteomic analysis of Neisseria gonorrhoeae biofilms shows shift to anaerobic respiration and changes in nutrient transport and outermembrane proteins

• Published in: PloS one 2012-06, Vol.7 (6), p.e38303-e38303
• Main Authors: Phillips, Nancy J, Steichen, Christopher T, Schilling, Birgit, Post, Deborah M B, Niles, Richard K, Bair, Thomas B, Falsetta, Megan L, Apicella, Michael A, Gibson, Bradford W
• Format: Article
• Language: English
• Subjects: Acinetobacter baumannii; Aging; Amino acids; Anaerobic respiration; Antibiotics; Archaea; Arginine; Bacteria; Bacteria, Anaerobic - growth & development; Bacteria, Anaerobic - metabolism; Bacterial Proteins - metabolism; Biofilms; Biofilms - growth & development; Biological Transport - genetics; Biology; Carbon Isotopes - metabolism; Cell culture; Chromatography; Chromatography, Liquid; Chronic infection; Cytochrome; Cytochrome c; Cytochrome-c Peroxidase - metabolism; Deoxyribonucleic acid; DNA; Drug dosages; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Energy metabolism; Flow chambers; Gel electrophoresis; Gels; Gene expression; Gene Expression Regulation, Bacterial - genetics; Gonorrhea; Health aspects; Immune system; Infections; Isotope Labeling; Liquid chromatography; Mass spectrometry; Mass spectroscopy; Medicine; Membrane proteins; Metabolism; Motility; Neisseria; Neisseria gonorrhoeae; Neisseria gonorrhoeae - growth & development; Neisseria gonorrhoeae - metabolism; Neisseria meningitidis; Nitrite reductase; Nitrite Reductases - metabolism; Nutrient transport; Organisms; Peroxidase; Planktonic cells; Protein biosynthesis; Protein expression; Protein synthesis; Protein turnover; Proteins; Proteomics; Proteomics - methods; Sexually transmitted diseases; Sodium lauryl sulfate; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Stable isotopes; Staphylococcus aureus; STD; Tandem Mass Spectrometry; Transcription
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0038303

Neisseria gonorrhoeae, the causative agent of gonorrhea, can form biofilms in vitro and in vivo. In biofilms, the organism is more resistant to antibiotic treatment and can serve as a reservoir for chronic infection. We have used stable isotope labeling by amino acids in cell culture (SILAC) to compare protein expression in biofilm and planktonic organisms. Two parallel populations of N. gonorrhoeae strain 1291, which is an arginine auxotroph, were grown for 48 h in continuous-flow chambers over glass, one supplemented with (13)C(6)-arginine for planktonic organisms and the other with unlabeled arginine for biofilm growth. The biofilm and planktonic cells were harvested and lysed separately, and fractionated into three sequential protein extracts. Corresponding heavy (H) planktonic and light (L) biofilm protein extracts were mixed and separated by 1D SDS-PAGE gels, and samples were extensively analyzed by liquid chromatography-mass spectrometry. Overall, 757 proteins were identified, and 152 unique proteins met a 1.5-fold cutoff threshold for differential expression with p-values