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

Xia Wu, Richard J. Siehnel, Jayanthi Garudathri, Benjamin J. Staudinger, Katherine B. Hisert, Egon Anderson Ozer, Alan R Hauser, Jimmy K. Eng, Colin Manoil, Pradeep K. Singh, James E. Bruce*

*Corresponding author for this work

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)2601-2612
Number of pages12
JournalJournal of Proteome Research
Volume18
Issue number6
DOIs
StatePublished - Jun 7 2019

Fingerprint

Proteome
Cystic Fibrosis
Pseudomonas aeruginosa
Physiology
Sputum
Siderophores
Centrifugation
Metabolism
Detergents
Labels
Lactic Acid
Bacteria
Carbon
Anti-Bacterial Agents
Peptides
Up-Regulation
Therapeutics
Proteins
Infection

Keywords

  • Pseudomonas aeruginosa
  • bacterial population proteomes
  • cystic fibrosis
  • quantitative proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Wu, X., Siehnel, R. J., Garudathri, J., Staudinger, B. J., Hisert, K. B., Ozer, E. A., ... Bruce, J. E. (2019). In Vivo Proteome of Pseudomonas aeruginosa in Airways of Cystic Fibrosis Patients. Journal of Proteome Research, 18(6), 2601-2612. https://doi.org/10.1021/acs.jproteome.9b00122
Wu, Xia ; Siehnel, Richard J. ; Garudathri, Jayanthi ; Staudinger, Benjamin J. ; Hisert, Katherine B. ; Ozer, Egon Anderson ; Hauser, Alan R ; Eng, Jimmy K. ; Manoil, Colin ; Singh, Pradeep K. ; Bruce, James E. / In Vivo Proteome of Pseudomonas aeruginosa in Airways of Cystic Fibrosis Patients. In: Journal of Proteome Research. 2019 ; Vol. 18, No. 6. pp. 2601-2612.
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abstract = "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.",
keywords = "Pseudomonas aeruginosa, bacterial population proteomes, cystic fibrosis, quantitative proteomics",
author = "Xia Wu and Siehnel, {Richard J.} and Jayanthi Garudathri and Staudinger, {Benjamin J.} and Hisert, {Katherine B.} and Ozer, {Egon Anderson} and Hauser, {Alan R} and Eng, {Jimmy K.} and Colin Manoil and Singh, {Pradeep K.} and Bruce, {James E.}",
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Wu, X, Siehnel, RJ, Garudathri, J, Staudinger, BJ, Hisert, KB, Ozer, EA, Hauser, AR, Eng, JK, Manoil, C, Singh, PK & Bruce, JE 2019, 'In Vivo Proteome of Pseudomonas aeruginosa in Airways of Cystic Fibrosis Patients', Journal of Proteome Research, vol. 18, no. 6, pp. 2601-2612. https://doi.org/10.1021/acs.jproteome.9b00122

In Vivo Proteome of Pseudomonas aeruginosa in Airways of Cystic Fibrosis Patients. / Wu, Xia; Siehnel, Richard J.; Garudathri, Jayanthi; Staudinger, Benjamin J.; Hisert, Katherine B.; Ozer, Egon Anderson; Hauser, Alan R; Eng, Jimmy K.; Manoil, Colin; Singh, Pradeep K.; Bruce, James E.

In: Journal of Proteome Research, Vol. 18, No. 6, 07.06.2019, p. 2601-2612.

Research output: Contribution to journalArticle

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T1 - In Vivo Proteome of Pseudomonas aeruginosa in Airways of Cystic Fibrosis Patients

AU - Wu, Xia

AU - Siehnel, Richard J.

AU - Garudathri, Jayanthi

AU - Staudinger, Benjamin J.

AU - Hisert, Katherine B.

AU - Ozer, Egon Anderson

AU - Hauser, Alan R

AU - Eng, Jimmy K.

AU - Manoil, Colin

AU - Singh, Pradeep K.

AU - Bruce, James E.

PY - 2019/6/7

Y1 - 2019/6/7

N2 - 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.

AB - 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.

KW - Pseudomonas aeruginosa

KW - bacterial population proteomes

KW - cystic fibrosis

KW - quantitative proteomics

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