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Literature summary for 1.97.1.4 extracted from

  • Crain, A.; Broderick, J.
    Pyruvate formate-lyase and its activation by pyruvate formate-lyase activating enzyme (2014), J. Biol. Chem., 289, 5723-5729 .
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
pyruvate pyruvate or oxamate are required for optimal activation Escherichia coli
oxamate pyruvate or oxamate are required for optimal activation Escherichia coli

Engineering

Protein Variants Comment Organism
additional information for activity assay, the enzyme PFL-AE is attached to a CM5 sensor chip using standard thiol coupling procedures Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics and equilibrium constant for the enzyme's interaction with substrate PFL, the interaction is very slow and rate-limited by large conformational changes, circular dichroism study Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
[4Fe-4S] cluster the [4Fe-4S] cluster of enzyme PFL-AE is coordinated by the cysteines of a conserved CX3CX2C motif, with the fourth unique iron coordinated by S-adenosyl-L-methionine. PFL-AE contains six cysteine residues (Cys12, Cys29, Cys33, Cys36, Cys94, Cys102) and only Cys29, Cys33, and Cys36 are involved in coordinating the iron sulfur cluster Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Escherichia coli in vivo concentrations of the entire PFL system is calculated to estimate the amount of bound protein in the cell. PFL, PFL-AE, and S-adenosyl-L-methionine are essentially fully bound in vivo, whereas electron donor proteins are partially bound ?
-
?
S-adenosyl-L-methionine + dihydroflavodoxin + [formate C-acetyltransferase]-glycine Escherichia coli
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5'-deoxyadenosine + L-methionine + flavodoxin semiquinone + [formate C-acetyltransferase]-glycin-2-yl radical
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli P0A9N4
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information in vivo concentrations of the entire PFL system is calculated to estimate the amount of bound protein in the cell. PFL, PFL-AE, and S-adenosyl-L-methionine are essentially fully bound in vivo, whereas electron donor proteins are partially bound Escherichia coli ?
-
?
additional information usage of an S-adenosyl-L-methionine binding assay to accurately determine the equilibrium constants for S-adenosyl-L-methionine binding to enzyme PFL-AE alone and in complex with substrate PFL, activation of PFL in the presence of its substrate pyruvate or the analogue oxamate results in stoichiometric conversion of the [4Fe-4S]1+ cluster to the glycyl radical on PFL. 3.7fold less activation is achieved in the absence of these small molecules, demonstrating that pyruvate or oxamate are required for optimal activation. For the assay, the enzyme PFL-AE is attached to a CM5 sensor chip using standard thiol coupling procedures Escherichia coli ?
-
?
S-adenosyl-L-methionine + dihydroflavodoxin + [formate C-acetyltransferase]-glycine
-
Escherichia coli 5'-deoxyadenosine + L-methionine + flavodoxin semiquinone + [formate C-acetyltransferase]-glycin-2-yl radical
-
?
S-adenosyl-L-methionine + dihydroflavodoxin + [formate C-acetyltransferase]-glycine PFL dimer, photoreduced enzyme PFL-AE Escherichia coli 5'-deoxyadenosine + L-methionine + flavodoxin semiquinone + [formate C-acetyltransferase]-glycin-2-yl radical
-
?

Synonyms

Synonyms Comment Organism
Pyruvate formate-lyase activating enzyme
-
Escherichia coli
PFL-AE
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Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
S-adenosyl-L-methionine
-
Escherichia coli

General Information

General Information Comment Organism
physiological function PFL-AE is a radical S-adenosyl-L-methionine enzyme that utilizes an iron-sulfur cluster and S-adenosyl-L-methionine to activate pyruvate formate lyase (PFL) via pro-S hydrogen abstraction from Gly734 Escherichia coli