EC Number   |
Activating Compound |
Reference |
|---|
    2.3.1.54 | pyruvate formate-lyase activating enzyme |
PFL-AE, EC 1.97.1.4, 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. 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. PFL activation studies, binding kinetics, overview |
745316 |
| 2.3.1.54 | pyruvate formate-lyase activating enzyme |
PFL-AE, EC 1.97.1.4, pyruvate formate-lyase is a glycyl radical enzyme activated by a radical AdoMet-activating enzyme PFL-AE, that exists largely in complex with enzyme PFL and S-adenosyl-L-methionine, all fully bound. Activation of pyruvate formate-lyase by pyruvate formate-lyase activating enzyme involves formation of a specific glycyl radical on pyruvate formate-lyase by the pyruvate formate-lyase activating enzyme in a reaction requiring S-adenosyl-L-methionine. 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 PF, 3.7fold less activation is achieved in the absence of these small molecules, demonstrating that pyruvate or oxamate are required for optimal activation. The [4Fe-4S] cluster of 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 cycles between two different oxidation states during hydrogen abstraction, [4Fe-4S]2+/1+, with the [4Fe-4S]1+ being the state capable of reductive cleavage of AdoMet to generate the glycyl radical |
727989 |
| 2.3.1.54 | pyruvate formate-lyase activating enzyme |
PflA, the enzyme is required to convert pyruvate formate-lyase, PflB, from its inactive to its active, glycyl-radical-containing species |
736638 |
| 2.3.1.54 | reduced ferredoxin |
reduced flavodoxin is te better cofactor |
736128 |
| 2.3.1.54 | reduced flavodoxin |
a better electron donor than reduced ferredoxin |
736128 |
| 2.3.1.54 | S-adenosylmethionine |
- |
14936, 487219, 487222, 487230 |
| 2.3.1.54 | S-adenosylmethionine |
obligatory component of the activation reaction |
487218, 487221, 487239 |
| 2.3.1.54 | Y06I |
autonomous glycyl radical cofactor, reconstituting the catalytic center of oxygen-fragmented enzyme |
487235 |
| 2.3.1.54 | YfiD |
- |
487235 |
| 2.3.1.54 | YfiD |
autonomous glycyl radical cofactor, reconstituting the catalytic center of oxygen-fragmented enzyme |
487235 |
