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  4. Absolute quantification of pceABCT gene products and characterization of the membrane protein complex responsible for the reduction of tetrachloroethene in Dehalobacter restrictus
 
conference presentation

Absolute quantification of pceABCT gene products and characterization of the membrane protein complex responsible for the reduction of tetrachloroethene in Dehalobacter restrictus

Cimmino, Lorenzo  
•
Duarte, Americo W.
•
Schmid, Adrian
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September 27, 2021
DehaloCon III 3rd International Conference on Anaerobic Biological Dehalogenation

The pceABCT gene cluster from Dehalobacter restrictus is one model system for tetrachloroethene (PCE) respiration. In the present study we aim to decipher the stoichiometry of the pceABCT gene products and the composition of PceA-containing membrane-bound protein complexes in D. restrictus. Quantitative proteomics (qProt), membrane extraction, CN-PAGE, and an in-gel enzymatic assay were developed. A qProt analysis performed on the membrane fraction revealed a 1.5:1 stoichiometric relationship between PceA and PceB, while PceC was about 60-times less abundant than PceA. Data from CN-PAGE and a newly developed in-gel PceA enzymatic assay revealed several forms of active PceA-containing complexes with masses of approximately 140, 180, and 250 kDa, depending on the physiological state of D. restrictus cells.

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Type
conference presentation
Author(s)
Cimmino, Lorenzo  
Duarte, Americo W.
Schmid, Adrian
Pereira, Ines A.C.
Holliger, Christof  
Maillard, Julien  
Date Issued

2021-09-27

Subjects

Dehalobacter restrictus

•

reductive dehalogenase complex

•

quantitative proteomics

Written at

EPFL

EPFL units
LBE  
Event nameEvent placeEvent date
DehaloCon III 3rd International Conference on Anaerobic Biological Dehalogenation

online

September 27-30, 2021

Available on Infoscience
February 1, 2022
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/185104
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