Loading…
Identification of a multi‐protein reductive dehalogenase complex in D ehalococcoides mccartyi strain CBDB 1 suggests a protein‐dependent respiratory electron transport chain obviating quinone involvement
Summary D ehalococcoides mccartyi strain CBDB 1 is an obligate organohalide‐respiring bacterium using only hydrogen as electron donor and halogenated organics as electron acceptor. Here, we studied proteins involved in the respiratory chain under non‐denaturing conditions. Using blue native gel elec...
Saved in:
Published in: | Environmental microbiology 2016-09, Vol.18 (9), p.3044-3056 |
---|---|
Main Authors: | , , , , , , , |
Format: | Article |
Language: | English |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
Summary: | Summary
D
ehalococcoides mccartyi
strain
CBDB
1 is an obligate organohalide‐respiring bacterium using only hydrogen as electron donor and halogenated organics as electron acceptor. Here, we studied proteins involved in the respiratory chain under non‐denaturing conditions. Using blue native gel electrophoresis (
BN‐PAGE
), gel filtration and ultrafiltration an active dehalogenating protein complex with a molecular mass of 250–270 kDa was identified. The active subunit of reductive dehalogenase (
RdhA
) colocalised with a complex iron‐sulfur molybdoenzyme (
CISM
) subunit (
CbdbA
195) and an iron‐sulfur cluster containing subunit (
CbdbA
131) of the hydrogen uptake hydrogenase (Hup). No colocalisation between the catalytically active subunits of hydrogenase and reductive dehalogenase was found. By two‐dimensional
BN
/
SDS‐PAGE
the stability of the complex towards detergents was assessed, demonstrating stepwise disintegration with increasing detergent concentrations. Chemical cross‐linking confirmed the presence of a higher molecular mass reductive dehalogenase protein complex composed of
RdhA
,
CISM I
and Hup hydrogenase and proved to be a potential tool for stabilising protein–protein interactions of the dehalogenating complex prior to membrane solubilisation. Taken together, the identification of the respiratory dehalogenase protein complex and the absence of indications for quinone participation in the respiration suggest a quinone‐independent protein‐based respiratory electron transfer chain in
D
. mccartyi
. |
---|---|
ISSN: | 1462-2912 1462-2920 |
DOI: | 10.1111/1462-2920.13200 |