2.3.1.54	Ferredoxin	-	487218, 487219, 487222	
2.3.1.54	Ferredoxin	methylviologen can replace ferredoxin	487222	
2.3.1.54	Ferredoxin	reactivation of inactive enzyme requires reduced ferredoxin, Fe2+-dithiol or Co2+-mercaptoethanol complexes can substitute for ferredoxin	487218	
2.3.1.54	additional information	conversion into the active form is carried out by an activating system containing a flavodoxin system and an activating enzyme (pyruvate format lyase-activase, EC 1.97.1.4)	663000	
2.3.1.54	NADPH	more effective for activation than NADPH, 0.02 mM NADPH activates over 80% inactive enzyme in cell-free extracts	663000	
2.3.1.54	oxamate	-	487222	
2.3.1.54	oxamate	obligatory component of the activation reaction	487221, 487239	
2.3.1.54	pyruvate	obligatory component of the activation reaction	487221, 487239	
2.3.1.54	pyruvate formate lyase activating enzyme	activates the enzyme by forming a glycine radical, cf. EC 1.97.1.4	744943	
2.3.1.54	pyruvate formate-lyase activating enzyme	converts the enzyme to its catalytically active state by generating a stable glycyl radical in the active site at G734	757586	
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