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  • 1
    Online Resource
    Online Resource
    Wiley ; 2016
    In:  PROTEOMICS – Clinical Applications Vol. 10, No. 9-10 ( 2016-10), p. 1036-1048
    In: PROTEOMICS – Clinical Applications, Wiley, Vol. 10, No. 9-10 ( 2016-10), p. 1036-1048
    Abstract: Trimethoprim is a folate biosynthesis inhibitor. Tetrahydrofolates are essential for the transfer of C 1 units in several biochemical pathways including purine, thymine, methionine, and glycine biosynthesis. This study addressed the effects of folate biosynthesis inhibition on bacterial physiology. Experimental design Two complementary proteomic approaches were employed to analyze the response of Bacillus subtilis to trimethoprim. Acute changes in protein synthesis rates were monitored by radioactive pulse labeling of newly synthesized proteins and subsequent 2DE analysis. Changes in protein levels were detected using gel‐free quantitative MS. Results Proteins involved in purine and histidine biosynthesis, the σ B ‐dependent general stress response, and sporulation were upregulated. Most prominently, the PurR‐regulon required for de novo purine biosynthesis was derepressed indicating purine depletion. The general stress response was activated energy dependently and in a subpopulation of treated cultures an early onset of sporulation was observed, most likely triggered by low guanosine triphosphate levels. Supplementation of adenosine triphosphate, adenosine, and guanosine to the medium substantially decreased antibacterial activity, showing that purine depletion becomes the bottleneck in trimethoprim‐treated B. subtilis . Conclusions and clinical relevance The frequently prescribed antibiotic trimethoprim causes purine depletion in B. subtilis , which can be complemented by supplementing purines to the medium.
    Type of Medium: Online Resource
    ISSN: 1862-8346 , 1862-8354
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2016
    detail.hit.zdb_id: 2317130-3
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  • 2
    In: PROTEOMICS, Wiley, Vol. 21, No. 1 ( 2021-01)
    Abstract: Identification of the molecular target is a crucial step in evaluating novel antibiotics. To support target identification, a label‐free method based on chromatographic co‐elution has previously been developed. Target identification by chromatographic coelution (TICC) exploits the alteration of the elution profile of target‐bound drug versus free drug in ion exchange (IEX) chromatography to identify potential target proteins from elution fractions. The applicability of TICC for antibiotic research is investigated by evaluating which proteins, that is, putative targets, can be monitored in Bacillus subtilis . Coelution of components of known protein complexes provides a read‐out for how well the native state of proteins is conserved during chromatography. Rifampicin, which targets RNA polymerase, is used in a proof‐of‐concept study.
    Type of Medium: Online Resource
    ISSN: 1615-9853 , 1615-9861
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2021
    detail.hit.zdb_id: 2037674-1
    SSG: 12
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  • 3
    In: MicrobiologyOpen, Wiley, Vol. 8, No. 12 ( 2019-12)
    Abstract: Rhodobacter capsulatus fixes atmospheric nitrogen (N 2 ) by a molybdenum (Mo)‐nitrogenase and a Mo‐free iron (Fe)‐nitrogenase, whose production is induced or repressed by Mo, respectively. At low nanomolar Mo concentrations, both isoenzymes are synthesized and contribute to nitrogen fixation. Here we examined the regulatory interplay of the central transcriptional activators NifA and AnfA by proteome profiling. As expected from earlier studies, synthesis of the structural proteins of Mo‐nitrogenase (NifHDK) and Fe‐nitrogenase (AnfHDGK) required NifA and AnfA, respectively, both of which depend on the alternative sigma factor RpoN to activate expression of their target genes. Unexpectedly, NifA was found to be essential for the synthesis of Fe‐nitrogenase, electron supply to both nitrogenases, biosynthesis of their cofactors, and production of RpoN. Apparently, RpoN is the only NifA‐dependent factor required for target gene activation by AnfA, since plasmid‐borne rpoN restored anfH transcription in a NifA‐deficient strain. However, plasmid‐borne rpoN did not restore Fe‐nitrogenase activity in this strain. Taken together, NifA requirement for synthesis and activity of both nitrogenases suggests that Fe‐nitrogenase functions as a complementary nitrogenase rather than an alternative isoenzyme in R .  capsulatus .
    Type of Medium: Online Resource
    ISSN: 2045-8827 , 2045-8827
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2019
    detail.hit.zdb_id: 2661368-2
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