A radical-chemical route to acetyl-CoA: the anaerobically induced pyruvate formate-lyase system of Escherichia coli.
Anaerobically growing Escherichia coli cells contain the enzyme pyruvate formate-lyase which catalyses the non-oxidative cleavage of pyruvate to acetyl-CoA and formate. The enzyme is subject to interconversion between inactive and active forms. The active form contains an oxygen-sensitive organic free radical located on the polypeptide chain which is essential for catalysis. It affords a novel homolytic C-C bond cleavage of the pyruvate substrate. The radical is generated by an iron-dependent converter enzyme which requires reduced flavodoxin and adenosyl methionine as co-substrates and pyruvate as a positive allosteric effector. A second converter enzyme, also iron-dependent, accomplishes the removal of the radical. This post-translational interconversion cycle controls the activity state of pyruvate formate-lyase in the anaerobic cell. Anaerobic conditions also regulate pyruvate formate-lyase at the level of gene expression. Multiple promoters are responsible for effecting a twelve to fifteen fold induction and they are coordinately controlled in response to the oxygen and metabolic status of the cell by sequences which are located far upstream of the pfl coding region. The transcription factor Fnr has been identified as being responsible for part of the anaerobic control of pfl expression, probably through direct interaction with the upstream sequences. In contrast, the expression of the gene encoding the first iron-dependent converter enzyme is unaffected by anaerobiosis and is independent of the Fnr protein.
Knappe
J
J
Institut für Biologische Chemie, Universität Heidelberg, F.R.G.
Sawers
G
G
D000105:Acetyl Coenzyme A / Q000096:biosynthesis*; D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism*; D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D016384:Consensus Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins; D005418:Flavodoxin / Q000378:metabolism; D005561:Formates / Q000378:metabolism; D005609:Free Radicals; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005809:Genes, Regulator; D007506:Iron-Sulfur Proteins; D008969:Molecular Sequence Data; D011773:Pyruvates / Q000378:metabolism; D019289:Pyruvic Acid; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C060704:FNR protein, E coli; D005418:Flavodoxin; D005561:Formates; D005609:Free Radicals; D007506:Iron-Sulfur Proteins; D011773:Pyruvates; C030544:formic acid; D000105:Acetyl Coenzyme A; D019289:Pyruvic Acid; D000123:Acetyltransferases; C022560:formate C-acetyltransferase