Regulation of substrate dependent expression and activity from dehalogenating enzymes in Dehalococcoides mccartyi strain CBDB1
Final Report Abstract
Wichtigste Ergebnisse: Thermal proteome profiling (TPP) was modified for analyzing bacterial oxygen-sensitive enzymes. Protein-trichloroethene interactions in organohalide-respiring Sulfurospirillum multivorans were identified. Interaction of the tetrachloroethene reductive dehalogenase with trichloroethene was confirmed. A first hint for the interaction of a response regulator and a chlorinated ethene was provided. - First integrated omics approach including proteomics and acetylomics of an Epsilonproteobacterium. Systematic comparison of protein production and regulatory mechanisms in organohalide respiration. Improved protein extraction allowed higher coverage of membrane proteins. Lysine acetylations within the two-component regulatory system might be involved in long-term downregulation. - First reference proteome of Dehalococcoides mccartyi strain CBDB1. Identification of 8491 distinct peptides which represent 1023 proteins that cover 70% of all annotated proteins in strain CBDB1. Spectra library was generated to allow fast and precise protein identifications for the scientific community. A proteogenomics workflow led to a refinement of gene annotation of CBDB1 and identified 4 new to date not annotated gene sequences. - An optimal quantification assay for proteins (namely selective reaction monitoring (SRM) and precursor reaction monitoring (PRM)) with the focus on those involved in housekeeping functions and putative reductive dehalogenase proteins from the strictly anaerobic bacterium Dehalococcoides mccartyi strain CBDB1 was established.
Publications
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Comparison of targeted peptide quantification assays for reductive dehalogenases by selective reaction monitoring (SRM) and precursor reaction monitoring (PRM). Anal Bioanal Chem. 2014 Jan;406(1):283-91
Schiffmann C, Hansen R, Baumann S, Kublik A, Nielsen PH, Adrian L, von Bergen M, Jehmlich N, Seifert J
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Proteome profile and proteogenomics of the organohalide-respiring bacterium Dehalococcoides mccartyi strain CBDB1 grown on hexachlorobenzene as electron acceptor. J Proteomics. 2014 Feb 26;98:59-64
Schiffmann CL, Jehmlich N, Otto W, Hansen R, Nielsen PH, Adrian L, Seifert J, von Bergen M
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Proteomic data set of the organohalide-respiring Epsilonproteobacterium Sulfurospirillum multivorans adapted to tetrachloroethene and other energy substrates. Data Brief. 2016 Jun 21;8:637-42
Goris T, Schiffmann CL, Gadkari J, Adrian L, von Bergen M, Diekert G, Jehmlich N
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Proteomic dataset of the organohalide-respiring bacterium Dehalococcoides mccartyi strain CBDB1 grown on hexachlorobenzene as electron acceptor. Data Brief. 2016 Feb 22;7:253-6
Schiffmann CL, Otto W, Hansen R, Nielsen PH, Adrian L, Seifert J, von Bergen M, Jehmlich N
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A retentive memory of tetrachloroethene respiration in Sulfurospirillum halorespirans - involvedp and a possible link to acetylation of a two-component regulatory system. J Proteomics. 2018 Jun 15;181:36-46
Türkowsky D, Esken J, Goris T, Schubert T, Diekert G, Jehmlich N, von Bergen M
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Thermal proteome profiling allows quantitative assessment of interactions between tetrachloroethene reductive dehalogenase and trichloroethene. J Proteomics. 2019 Feb 10;192:10-17
Türkowsky D, Lohmann P, Mühlenbrink M, Schubert T, Adrian L, Goris T, Jehmlich N, von Bergen M
