Proteomic profiling of Mycobacterium tuberculosis identifies nutrient-starvation-responsive toxin-antitoxin systems

Jakob Albrethsen, Jeppe Agner, Sander R Piersma, Peter Højrup, Thang V Pham, Karin Weldingh, Connie R Jimenez, Peter Andersen, Ida Rosenkrands

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

In order to successfully enter the latent stage, Mycobacterium tuberculosis must adapt to conditions such as nutrient limitation and hypoxia. In vitro models that mimic latent infection are valuable tools for describing the changes in metabolism that occur when the bacterium exists in a non-growing form. We used two complementary proteomic approaches, label-free LC-MS/MS analysis and two-dimensional difference gel electrophoresis, to determine the proteome profile of extracellular proteins from M. tuberculosis cultured under nutrient starvation. Through the label-free LC-MS/MS analysis of fractionated samples, 1176 proteins were identified from culture filtrates of log phase and nutrient-starved cultures, and the protein levels of 230 proteins were increased in nutrient-starved culture filtrates, whereas those of 208 proteins were decreased. By means of Gene Ontology clustering analysis, significant differences in the overall metabolism during nutrient starvation were detected. Notably, members of the toxin-antitoxin systems were present in larger quantities in nutrient-starved cultures, supporting a role for these global modules as M. tuberculosis switches its metabolism into dormancy. Decreased abundance of proteins involved in amino acid and protein synthesis was apparent, as well as changes in the lipid metabolism. Further analysis of the dataset identified increased abundance of lipoproteins and decreased abundance of ESAT-6 family proteins. Results from the two-dimensional difference gel electrophoresis proteomics demonstrated overall agreement with the LC-MS/MS data and added complementary insights about protein degradation and modification.
Original languageEnglish
JournalMolecular and Cellular Proteomics
Volume12
Issue number5
Pages (from-to)1180-1191
ISSN1535-9476
DOIs
Publication statusPublished - 2013

Fingerprint

Antitoxins
Mycobacterium tuberculosis
Nutrients
Food
Proteins
Two-Dimensional Difference Gel Electrophoresis
Metabolism
Electrophoresis
Labels
Gels
Gene Ontology
Proteomics
Proteome
Lipid Metabolism
Proteolysis
Lipoproteins
Cluster Analysis
Ontology
Bacteria
Genes

Cite this

Albrethsen, Jakob ; Agner, Jeppe ; Piersma, Sander R ; Højrup, Peter ; Pham, Thang V ; Weldingh, Karin ; Jimenez, Connie R ; Andersen, Peter ; Rosenkrands, Ida. / Proteomic profiling of Mycobacterium tuberculosis identifies nutrient-starvation-responsive toxin-antitoxin systems. In: Molecular and Cellular Proteomics. 2013 ; Vol. 12, No. 5. pp. 1180-1191.
@article{0488bdb55d9b4d6aa766b3e527419893,
title = "Proteomic profiling of Mycobacterium tuberculosis identifies nutrient-starvation-responsive toxin-antitoxin systems",
abstract = "In order to successfully enter the latent stage, Mycobacterium tuberculosis must adapt to conditions such as nutrient limitation and hypoxia. In vitro models that mimic latent infection are valuable tools for describing the changes in metabolism that occur when the bacterium exists in a non-growing form. We used two complementary proteomic approaches, label-free LC-MS/MS analysis and two-dimensional difference gel electrophoresis, to determine the proteome profile of extracellular proteins from M. tuberculosis cultured under nutrient starvation. Through the label-free LC-MS/MS analysis of fractionated samples, 1176 proteins were identified from culture filtrates of log phase and nutrient-starved cultures, and the protein levels of 230 proteins were increased in nutrient-starved culture filtrates, whereas those of 208 proteins were decreased. By means of Gene Ontology clustering analysis, significant differences in the overall metabolism during nutrient starvation were detected. Notably, members of the toxin-antitoxin systems were present in larger quantities in nutrient-starved cultures, supporting a role for these global modules as M. tuberculosis switches its metabolism into dormancy. Decreased abundance of proteins involved in amino acid and protein synthesis was apparent, as well as changes in the lipid metabolism. Further analysis of the dataset identified increased abundance of lipoproteins and decreased abundance of ESAT-6 family proteins. Results from the two-dimensional difference gel electrophoresis proteomics demonstrated overall agreement with the LC-MS/MS data and added complementary insights about protein degradation and modification.",
author = "Jakob Albrethsen and Jeppe Agner and Piersma, {Sander R} and Peter H{\o}jrup and Pham, {Thang V} and Karin Weldingh and Jimenez, {Connie R} and Peter Andersen and Ida Rosenkrands",
year = "2013",
doi = "10.1074/mcp.M112.018846",
language = "English",
volume = "12",
pages = "1180--1191",
journal = "Molecular and Cellular Proteomics",
issn = "1535-9476",
publisher = "American Society for Biochemistry and Molecular Biology",
number = "5",

}

Albrethsen, J, Agner, J, Piersma, SR, Højrup, P, Pham, TV, Weldingh, K, Jimenez, CR, Andersen, P & Rosenkrands, I 2013, 'Proteomic profiling of Mycobacterium tuberculosis identifies nutrient-starvation-responsive toxin-antitoxin systems', Molecular and Cellular Proteomics, vol. 12, no. 5, pp. 1180-1191. https://doi.org/10.1074/mcp.M112.018846

Proteomic profiling of Mycobacterium tuberculosis identifies nutrient-starvation-responsive toxin-antitoxin systems. / Albrethsen, Jakob; Agner, Jeppe; Piersma, Sander R; Højrup, Peter; Pham, Thang V; Weldingh, Karin; Jimenez, Connie R; Andersen, Peter; Rosenkrands, Ida.

In: Molecular and Cellular Proteomics, Vol. 12, No. 5, 2013, p. 1180-1191.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Proteomic profiling of Mycobacterium tuberculosis identifies nutrient-starvation-responsive toxin-antitoxin systems

AU - Albrethsen, Jakob

AU - Agner, Jeppe

AU - Piersma, Sander R

AU - Højrup, Peter

AU - Pham, Thang V

AU - Weldingh, Karin

AU - Jimenez, Connie R

AU - Andersen, Peter

AU - Rosenkrands, Ida

PY - 2013

Y1 - 2013

N2 - In order to successfully enter the latent stage, Mycobacterium tuberculosis must adapt to conditions such as nutrient limitation and hypoxia. In vitro models that mimic latent infection are valuable tools for describing the changes in metabolism that occur when the bacterium exists in a non-growing form. We used two complementary proteomic approaches, label-free LC-MS/MS analysis and two-dimensional difference gel electrophoresis, to determine the proteome profile of extracellular proteins from M. tuberculosis cultured under nutrient starvation. Through the label-free LC-MS/MS analysis of fractionated samples, 1176 proteins were identified from culture filtrates of log phase and nutrient-starved cultures, and the protein levels of 230 proteins were increased in nutrient-starved culture filtrates, whereas those of 208 proteins were decreased. By means of Gene Ontology clustering analysis, significant differences in the overall metabolism during nutrient starvation were detected. Notably, members of the toxin-antitoxin systems were present in larger quantities in nutrient-starved cultures, supporting a role for these global modules as M. tuberculosis switches its metabolism into dormancy. Decreased abundance of proteins involved in amino acid and protein synthesis was apparent, as well as changes in the lipid metabolism. Further analysis of the dataset identified increased abundance of lipoproteins and decreased abundance of ESAT-6 family proteins. Results from the two-dimensional difference gel electrophoresis proteomics demonstrated overall agreement with the LC-MS/MS data and added complementary insights about protein degradation and modification.

AB - In order to successfully enter the latent stage, Mycobacterium tuberculosis must adapt to conditions such as nutrient limitation and hypoxia. In vitro models that mimic latent infection are valuable tools for describing the changes in metabolism that occur when the bacterium exists in a non-growing form. We used two complementary proteomic approaches, label-free LC-MS/MS analysis and two-dimensional difference gel electrophoresis, to determine the proteome profile of extracellular proteins from M. tuberculosis cultured under nutrient starvation. Through the label-free LC-MS/MS analysis of fractionated samples, 1176 proteins were identified from culture filtrates of log phase and nutrient-starved cultures, and the protein levels of 230 proteins were increased in nutrient-starved culture filtrates, whereas those of 208 proteins were decreased. By means of Gene Ontology clustering analysis, significant differences in the overall metabolism during nutrient starvation were detected. Notably, members of the toxin-antitoxin systems were present in larger quantities in nutrient-starved cultures, supporting a role for these global modules as M. tuberculosis switches its metabolism into dormancy. Decreased abundance of proteins involved in amino acid and protein synthesis was apparent, as well as changes in the lipid metabolism. Further analysis of the dataset identified increased abundance of lipoproteins and decreased abundance of ESAT-6 family proteins. Results from the two-dimensional difference gel electrophoresis proteomics demonstrated overall agreement with the LC-MS/MS data and added complementary insights about protein degradation and modification.

U2 - 10.1074/mcp.M112.018846

DO - 10.1074/mcp.M112.018846

M3 - Journal article

C2 - 23345537

VL - 12

SP - 1180

EP - 1191

JO - Molecular and Cellular Proteomics

JF - Molecular and Cellular Proteomics

SN - 1535-9476

IS - 5

ER -