title: High-Resolution Mass Spectrometry Based Proteomic Analysis of the Response to Vancomycin-Induced Cell Wall Stress in Streptomyces coelicolor A3(2).
abstract: Understanding how bacteria survive periods of cell wall stress is of fundamental interest and can help generate ideas for improved antibacterial treatments. In this study we use tandem mass tagging to characterize the proteomic response of vancomycin resistant Streptomyces coelicolor to the exposure to sublethal levels of the antibiotic. A common set of 804 proteins were identified in triplicate experiments. Contrasting changes in the abundance of proteins closely associated with the cytoplasmic membrane with those taking place in the cytosol identified aspects of protein spatial localization that are associated with the response to vancomycin. Enzymes for peptidoglycan precursor, mycothiol, ectoine and menaquinone biosynthesis together with a multisubunit nitrate reductase were recruited to the membrane following vancomycin treatment. Many proteins with regulatory functions (including sensor protein kinases) also exhibited significant changes in abundance exclusively in the membrane-associated protein fraction. Several enzymes predicted to be involved in extracellular peptidoglycan crossbridge formation became significantly depleted from the membrane. A comparison with data previously acquired on the changes in gene transcription following vancomycin treatment identified a common high-confidence set of changes in gene expression. Generalized changes in protein abundance indicate roles for proteolysis, the pentose phosphate pathway and a reorganization of amino acid biosynthesis in the stress response.
authors.affiliation: †Department of Biochemistry, University of Cambridge, Cambridge, U.K.|‡Cambridge Systems Biology Centre, University of Cambridge, Cambridge, U.K.
authors.forename: Andy
authors.initials: A
authors.lastname: Hesketh
authors.affiliation: †Department of Biochemistry, University of Cambridge, Cambridge, U.K.|‡Cambridge Systems Biology Centre, University of Cambridge, Cambridge, U.K.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Deery
authors.affiliation: †Department of Biochemistry, University of Cambridge, Cambridge, U.K.
authors.forename: Hee-Jeon
authors.initials: HJ
authors.lastname: Hong
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000378:metabolism*; D002473:Cell Wall / Q000187:drug effects*; D002853:Chromatography, Liquid; D040901:Proteomics; D048372:Streptomyces coelicolor / Q000187:drug effects* / Q000378:metabolism; D013312:Stress, Physiological; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014640:Vancomycin / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D014640:Vancomycin
keywords: Streptomyces coelicolor; cell wall stress; proteome; transcriptome; vancomycin