In Vivo Proteome of Pseudomonas aeruginosa in Airways of Cystic Fibrosis Patients

• Published in: Journal of proteome research Vol. 18; no. 6; pp. 2601 - 2612
• Main Authors: Wu, Xia, Siehnel, Richard J, Garudathri, Jayanthi, Staudinger, Benjamin J, Hisert, Katherine B, Ozer, Egon A, Hauser, Alan R, Eng, Jimmy K, Manoil, Colin, Singh, Pradeep K, Bruce, James E
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 07.06.2019
• Subjects: Adult; Anti-Bacterial Agents - therapeutic use; Bacterial Proteins - genetics; Carbon - metabolism; Cystic Fibrosis - diagnosis; Cystic Fibrosis - drug therapy; Cystic Fibrosis - genetics; Cystic Fibrosis - microbiology; Female; Glyoxylates - metabolism; Humans; Lactic Acid - metabolism; Male; Membrane Proteins - genetics; Middle Aged; Proteome - genetics; Pseudomonas aeruginosa - drug effects; Pseudomonas aeruginosa - isolation & purification; Pseudomonas aeruginosa - pathogenicity; Pseudomonas Infections - diagnosis; Pseudomonas Infections - drug therapy; Pseudomonas Infections - genetics; Pseudomonas Infections - microbiology; Sputum - microbiology; Pseudomonas aeruginosa; cystic fibrosis; quantitative proteomics; bacterial population proteomes
• ISSN: 1535-3893; 1535-3907
• DOI: 10.1021/acs.jproteome.9b00122

Chronic airway infection with P. aeruginosa (PA) is a hallmark of cystic fibrosis (CF) disease. The mechanisms producing PA persistence in CF therapies remain poorly understood. To gain insight on PA physiology in patient airways and better understand how in vivo bacterial functioning differs from in vitro conditions, we investigated the in vivo proteomes of PA in 35 sputum samples from 11 CF patients. We developed a novel bacterial-enrichment method that relies on differential centrifugation and detergent treatment to enrich for bacteria to improve identification of PA proteome with CF sputum samples. Using two nonredundant peptides as a cutoff, a total of 1304 PA proteins were identified directly from CF sputum samples. The in vivo PA proteomes were compared with the proteomes of ex vivo-grown PA populations from the same patient sample. Label-free quantitation and proteome comparison revealed the in vivo up-regulation of siderophore TonB-dependent receptors, remodeling in central carbon metabolism including glyoxylate cycle and lactate utilization, and alginate overproduction. Knowledge of these in vivo proteome differences or others derived using the presented methodology could lead to future treatment strategies aimed at altering PA physiology in vivo to compromise infectivity or improve antibiotic efficacy.