Proteome assessment of organohalide respiring species and quantification of dehalogenating enzymes in pure and mixed cultures in response to various electron acceptors
Martin von Bergen / Nico Jehmlich (UFZ Leipzig)
High-resolution mass spectrometry techniques are used to obtain a deeper insight into the proteome of organohalid respiring bacteria and to determine the expression level of different dehalogenases. For the first objective shotgun proteomic analyses (LC-MS/MS) are performed to get a proteome inventory of different Dehalococcoides sp. and to elucidate the expression of biochemical pathways under different cultivation conditions. In the second objective selected reaction monitoring - mass spectrometry (srm-MS) is done to obtain targeted expression data of specific dehalogenases in the presence of different halogenated compounds. The specific quantification will yield insights into the correlation between substrate, culture conditions and involved biochemical pathways and the expression of orthologous reductive dehalogenases.
Scheme of shotgun proteomic analysis
Goris T, Schiffmann CL, Gadkari J, Schubert T, Seifert J, Jehmlich N, von Bergen M, Diekert G.
Proteomics of the organohalide-respiring Epsilonproteobacterium Sulfurospirillum multivorans adapted to tetrachloroethene and other energy substrates
Sci. Rep. 2015, (5)13794, Sep 21, doi:10.1038/srep13794
Schiffmann C.L., Jehmlich N., Otto W., Hansen R., Nielsen P.H., Adrian L., Seifert J., von Bergen M.
Proteome profile and proteogenomics of the organohalide-respiring bacterium Dehalococcoides mccartyi strain CBDB1 grown on hexachlorobenzene as electron acceptor
J Proteomics. 2013 Dec 25;98C:59-64 (Epub ahead of print) doi:10.1016/j.jprot.2013.12.009
Schiffmann C., Hansen R., Baumann S., Kublik A., Nielsen P.H., Adrian L., von Bergen M., Jehmlich N., Seifert J.
Comparison of targeted peptide quantification assays for reductive dehalogenases by selective reaction monitoring (SRM) and precursor reaction monitoring (PRM)
Anal Bioanal Chem. 2013 Nov 13. (Epub ahead of print) doi:10.1007/s00216-013-7451-7