Genome sequence, proteome profile, and identification of a multiprotein reductive dehalogenase complex in Dehalogenimonas alkenigignens strain BRE15M

Alba Trueba-Santiso, Kenneth Wasmund, Jesica M. Soder-Walz, Ernest Marco-Urrea*, Lorenz Adrian

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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Abstract

Bacteria of the genus Dehalogenimonas respire with vicinally halogenated alkanes via dihaloelimination. We aimed to describe involved proteins and their supermolecular organization. Metagenomic sequencing of a Dehalogenimonas-containing culture resulted in a 1.65 Mbp draft genome of Dehalogenimonas alkenigignens strain BRE15M. It contained 31 full-length reductive dehalogenase homologous genes (rdhA), but only eight had cognate rdhB gene coding for membrane-anchoring proteins. Shotgun proteomics of cells grown with 1,2-dichloropropane as an electron acceptor identified 1152 proteins representing more than 60% of the total proteome. Ten RdhA proteins were detected, including a DcpA ortholog, which was the strongest expressed RdhA. Blue native gel electrophoresis (BNE) demonstrating maximum activity was localized in a protein complex of 146-242 kDa. Protein mass spectrometry revealed the presence of DcpA, its membrane-anchoring protein DcpB, two hydrogen uptake hydrogenase subunits (HupL and HupS), an iron-sulfur protein (HupX), and subunits of a redox protein with a molybdopterin-binding motif (OmeA and OmeB) in the complex. BNE after protein solubilization with different detergent concentrations revealed no evidence for an interaction between the putative respiratory electron input module (HupLS) and the OmeA/OmeB/HupX module. All detected RdhAs comigrated with the organohalide respiration complex. Based on genomic and proteomic analysis, we propose quinone-independent respiration in Dehalogenimonas.

Original languageEnglish
Pages (from-to)613-623
Number of pages11
JournalJournal of Proteome Research
Volume20
Issue number1
Early online date25 Sep 2020
DOIs
Publication statusPublished - 1 Jan 2021

Keywords

  • 1,2-dichloropropane
  • Dehalococcoidia
  • Dehalogenimonas alkenigignens strain BRE15M
  • dihaloelimination
  • genome sequencing
  • organohalide respiration
  • proteome profiling

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