Literature summary for 2.5.1.7 extracted from

Kim, D.H.; Lees, W.J.; Kempsell, K.E.; Lane, W.S.; Duncan, K.; Walsh, C.T.
Characterization of a Cys115 to Asp substitution in the Escherichia coli cell wall biosynthetic enzyme UDP-GlcNAc enolpyruvyl transferase (MurA) that confers resistance to inactivation by the antibiotic fosfomycin (1996), Biochemistry, 35, 4923-4928.

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Protein Variants

Protein Variants	Comment	Organism
C115A	site-directed mutagenesis, overexpression in Escherichia coli, no activity	Escherichia coli
C115D	site-directed mutagenesis, overexpression in Escherichia coli, gains fosfomycin resistance, forms only the phospholactyl-enzyme intermediate adduct, but no UDP-GlcNAc-phosphoenolpyruvate, higher kcat than the wild-type at pH 7.0, enhanced pH-dependency of the reaction	Escherichia coli
C115E	site-directed mutagenesis, overexpression in Escherichia coli, gains fosfomycin resistance, enhanced pH-dependency of the reaction, low activity	Escherichia coli
C115N	site-directed mutagenesis, overexpression in Escherichia coli, deamination of Asn115 to Asp115	Escherichia coli
C115S	-	Enterobacter cloacae
C115S	site-directed mutagenesis, overexpression in Escherichia coli, no activity	Escherichia coli

Inhibitors

Inhibitors	Comment	Organism	Structure
(Z)-3-fluorophosphoenolpyruvate	kinetics, mutant C115D; wild-type and mutant C115D, competitive against phosphoenolpyruvate	Escherichia coli
fosfomycin	alkylates Cys115	Enterobacter cloacae
fosfomycin	alkylates Cys115; competitive against phosphoenolpyruvate; inhibits the wild-type, mutant C155D is completely resistant; t1/2 for inactivation of the wild-type enzyme: 6 s	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0004	-	phosphoenolpyruvate	wild-type enzyme	Escherichia coli
0.015	-	UDP-GlcNAc	wild-type enzyme	Escherichia coli
0.016	-	UDP-GlcNAc	mutant C115D, pH 8.0	Escherichia coli
0.021	-	UDP-GlcNAc	mutant C115D, pH 6.0	Escherichia coli
0.022	-	phosphoenolpyruvate	mutant C115D, pH 6.0	Escherichia coli
0.037	-	phosphoenolpyruvate	mutant C115D, pH 8.0	Escherichia coli

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining

Organism

Organism	UniProt	Comment	Textmining
Enterobacter cloacae	-	-	-
Escherichia coli	-	MurA	-
Escherichia coli MurA	-	MurA	-

Reaction

Reaction	Comment	Organism	Reaction ID
phosphoenolpyruvate + UDP-N-acetyl-alpha-D-glucosamine = phosphate + UDP-N-acetyl-3-O-(1-carboxyvinyl)-alpha-D-glucosamine	Cys115 is the active site nucleophile	Escherichia coli	a
phosphoenolpyruvate + UDP-N-acetyl-alpha-D-glucosamine = phosphate + UDP-N-acetyl-3-O-(1-carboxyvinyl)-alpha-D-glucosamine	Cys115 is the active site nucleophile	Enterobacter cloacae	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
phosphoenolpyruvate + UDP-N-acetyl-D-glucosamine	-	Escherichia coli	phosphate + UDP-N-acetyl-3-(1-carboxyvinyl)-D-glucosamine	i.e. UDP-N-acetyl-D-glucosamine-enolpyruvate	?
phosphoenolpyruvate + UDP-N-acetyl-D-glucosamine	-	Enterobacter cloacae	phosphate + UDP-N-acetyl-3-(1-carboxyvinyl)-D-glucosamine	i.e. UDP-N-acetyl-D-glucosamine-enolpyruvate	?
phosphoenolpyruvate + UDP-N-acetyl-D-glucosamine	-	Escherichia coli MurA	phosphate + UDP-N-acetyl-3-(1-carboxyvinyl)-D-glucosamine	i.e. UDP-N-acetyl-D-glucosamine-enolpyruvate	?

Synonyms

Synonyms	Comment	Organism
UDP-GlcNAc enolpyruvyl transferase	-	Escherichia coli
UDP-GlcNAc enolpyruvyl transferase	-	Enterobacter cloacae

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.0086	-	fosfomycin	wild-type enzyme	Escherichia coli
0.04	-	(Z)-3-fluorophosphoenolpyruvate	wild-type enzyme	Escherichia coli
0.04	-	(Z)-3-fluorophosphoenolpyruvate	at pH 8.0	Escherichia coli
0.08	-	(Z)-3-fluorophosphoenolpyruvate	at pH 8.0	Escherichia coli
0.08	-	(Z)-3-fluorophosphoenolpyruvate	mutant C115D	Escherichia coli
0.1	-	(Z)-3-fluorophosphoenolpyruvate	at pH 6.0	Escherichia coli
0.1	-	(Z)-3-fluorophosphoenolpyruvate	mutant C115D	Escherichia coli
1	-	fosfomycin	at pH 8.0	Escherichia coli
1	-	fosfomycin	mutant C115D	Escherichia coli
2	-	fosfomycin	at pH 6.0	Escherichia coli
2	-	fosfomycin	mutant C115D	Escherichia coli

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Release 2023.1 (January 2023)