Activating Compound | Comment | Organism | Structure |
---|---|---|---|
oxamate | pyruvate or oxamate are required for optimal activation | Escherichia coli | |
pyruvate | pyruvate or oxamate are required for optimal activation | Escherichia coli |
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 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 | 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 | 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 | - |
5'-deoxyadenosine + L-methionine + flavodoxin semiquinone + [formate C-acetyltransferase]-glycin-2-yl radical | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A9N4 | - |
- |
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 | Comment | Organism |
---|---|---|
PFL-AE | - |
Escherichia coli |
Pyruvate formate-lyase activating enzyme | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Escherichia coli |
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 |