The glycyl-radical enzyme 2-ketobutyrate formate-lyase, TdcE, interacts specifically with the formate-translocating FNT-channel protein FocA

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The glycyl-radical enzyme 2-ketobutyrate formate-lyase, TdcE, interacts specifically with the formate-translocating FNT-channel protein FocA

Formate is a major product of mixed-acid fermentation in Escherichia coli. Because formate can act as an uncoupler at high concentration it must be excreted from the cell. The FNT (formate-nitrite transporter) membrane channel FocA ensures formate is translocated across the cytoplasmic membrane. Two glycyl-radical enzymes (GREs), pyruvate formate-lyase (PflB) and 2-ketobutyrate formate-lyase (T...

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The glycyl radical enzyme TdcE can replace pyruvate formate-lyase in glucose fermentation.

Mutants of Escherichia coli unable to synthesize a functional pyruvate formate-lyase (PFL) are severely impaired in their capacity to grow by glucose fermentation. In a functional complementation assay designed to isolate the pfl gene from Clostridium butyricum, we fortuitously identified a gene that did not encode a PFL but nonetheless was able to complement the phenotypic defects caused by an...

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Pyruvate formate-lyase mechanism involving the protein-based glycyl radical.

Pyruvate formate-lyase (also called formate acetyltransferase; EC 2.3.1.54; PFI .) catalyses the thiolytic cleavage of pyruvate by CoA, yielding acetyl-CoA and formate. This reaction is the key step in the glucose-fermentation route in Escherichziz coli and various other bacteria. Operationally, it resembles the (B-keto)thiolase reaction of the fatty-acid degradation cycle. The mechanism of pyr...

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The C-terminal Six Amino Acids of the FNT Channel FocA Are Required for Formate Translocation But Not Homopentamer Integrity

FocA is the archetype of the pentameric formate-nitrite transporter (FNT) superfamily of channels, members of which translocate small organic and inorganic anions across the cytoplasmic membrane of microorganisms. The N- and C-termini of each protomer are cytoplasmically oriented. A Y-L-R motif is found immediately after transmembrane helix 6 at the C-terminus of FNT proteins related to FocA, o...

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Structure and mechanism of the glycyl radical enzyme pyruvate formate-lyase[6]

The enzyme pyruvate formate-lyase (PFL) catalyzes the reversible conversion of pyruvate and CoA into acetyl-CoA and formate, which has a central role in anaerobic glucose fermentation by E. coli cells and other bacteria [1]. PFL a 2 × 85 kDa homodimer is the first example of a radical enzyme where the spin was found to be located on the polypeptide backbone Cα-atom of a glycyl residue (Gly 734)...

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ژورنال

عنوان ژورنال: Biochemistry and Biophysics Reports

سال: 2016

ISSN: 2405-5808

DOI: 10.1016/j.bbrep.2016.04.005