Skip to Content
Merck
  • Top-down proteomics reveals a unique protein S-thiolation switch in Salmonella Typhimurium in response to infection-like conditions.

Top-down proteomics reveals a unique protein S-thiolation switch in Salmonella Typhimurium in response to infection-like conditions.

Proceedings of the National Academy of Sciences of the United States of America (2013-05-31)
Charles Ansong, Si Wu, Da Meng, Xiaowen Liu, Heather M Brewer, Brooke L Deatherage Kaiser, Ernesto S Nakayasu, John R Cort, Pavel Pevzner, Richard D Smith, Fred Heffron, Joshua N Adkins, Ljiljana Pasa-Tolic
ABSTRACT

Characterization of the mature protein complement in cells is crucial for a better understanding of cellular processes on a systems-wide scale. Toward this end, we used single-dimension ultra-high-pressure liquid chromatography mass spectrometry to investigate the comprehensive "intact" proteome of the Gram-negative bacterial pathogen Salmonella Typhimurium. Top-down proteomics analysis revealed 563 unique proteins including 1,665 proteoforms generated by posttranslational modifications (PTMs), representing the largest microbial top-down dataset reported to date. We confirmed many previously recognized aspects of Salmonella biology and bacterial PTMs, and our analysis also revealed several additional biological insights. Of particular interest was differential utilization of the protein S-thiolation forms S-glutathionylation and S-cysteinylation in response to infection-like conditions versus basal conditions. This finding of a S-glutathionylation-to-S-cysteinylation switch in a condition-specific manner was corroborated by bottom-up proteomics data and further by changes in corresponding biosynthetic pathways under infection-like conditions and during actual infection of host cells. This differential utilization highlights underlying metabolic mechanisms that modulate changes in cellular signaling, and represents a report of S-cysteinylation in Gram-negative bacteria. Additionally, the functional relevance of these PTMs was supported by protein structure and gene deletion analyses. The demonstrated utility of our simple proteome-wide intact protein level measurement strategy for gaining biological insight should promote broader adoption and applications of top-down proteomics approaches.

MATERIALS
Product Number
Brand
Product Description

Supelco
L-Cysteine hydrochloride monohydrate, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
L-Cysteine hydrochloride monohydrate, Produced by Wacker Chemie AG, Burghausen, Germany, Life Science, 98.5-101.0%
Sigma-Aldrich
L-Cysteine hydrochloride monohydrate, reagent grade, ≥98% (TLC)
Sigma-Aldrich
L-Cysteine hydrochloride monohydrate, from non-animal source, suitable for cell culture, meets EP, USP testing specifications
Sigma-Aldrich
L-Cysteine hydrochloride monohydrate, BioUltra, ≥99.0% (RT)
Cysteine hydrochloride monohydrate, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Sulfur, powder, 99.98% trace metals basis
Sigma-Aldrich
L-Cysteine, ≥97%, FG
Sigma-Aldrich
L-Cysteine, 97%
Supelco
Sulfur, PESTANAL®, analytical standard
Sigma-Aldrich
L-Cysteine, BioUltra, ≥98.5% (RT)
Sigma-Aldrich
Sulfur, flakes, ≥99.99% trace metals basis
Sigma-Aldrich
Sulfur, 99.998% trace metals basis
Sigma-Aldrich
L-Cysteine, from non-animal source, BioReagent, suitable for cell culture, ≥98%
Sigma-Aldrich
L-Cysteine, produced by Wacker Chemie AG, Burghausen, Germany, ≥98.0%
Supelco
L-Cysteine, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
SAFC
L-Cysteine
Sigma-Aldrich
L-Cysteine, Vetec, reagent grade, 97%