DB code: D00415

RLCP classification 4.23.18400.69 : Addition
5.303.667700.54 : Elimination
CATH domain 3.65.10.10 : UDP-n-acetylglucosamine1-carboxyvinyl-transferase; Chain Catalytic domain
3.65.10.10 : UDP-n-acetylglucosamine1-carboxyvinyl-transferase; Chain Catalytic domain
E.C. 2.5.1.7
CSA 1uae
M-CSA 1uae
MACiE

CATH domain Related DB codes (homologues)

Uniprot Enzyme Name
UniprotKB Protein name Synonyms RefSeq Pfam
P0A749 UDP-N-acetylglucosamine 1-carboxyvinyltransferase
EC 2.5.1.7
Enoylpyruvate transferase
UDP-N-acetylglucosamine enolpyruvyl transferase
EPT
NP_417656.1 (Protein)
NC_000913.2 (DNA/RNA sequence)
YP_491374.1 (Protein)
NC_007779.1 (DNA/RNA sequence)
PF00275 (EPSP_synthase)
[Graphical View]
P33038 UDP-N-acetylglucosamine 1-carboxyvinyltransferase
EC 2.5.1.7
Enoylpyruvate transferase
UDP-N-acetylglucosamine enolpyruvyl transferase
EPT
YP_003615048.1 (Protein)
NC_014121.1 (DNA/RNA sequence)
PF00275 (EPSP_synthase)
[Graphical View]

KEGG enzyme name
UDP-N-acetylglucosamine 1-carboxyvinyltransferase
MurA transferase
UDP-N-acetylglucosamine 1-carboxyvinyl-transferase
UDP-N-acetylglucosamine enoylpyruvyltransferase
enoylpyruvate transferase
phosphoenolpyruvate-UDP-acetylglucosamine-3-enolpyruvyltransferase
phosphoenolpyruvate:UDP-2-acetamido-2-deoxy-D-glucose2-enoyl-1-carboxyethyltransferase
phosphoenolpyruvate:uridine diphosphate N-acetylglucosamineenolpyruvyltransferase
phosphoenolpyruvate:uridine-5'-diphospho-N-acetyl-2-amino-2-deoxyglucose 3-enolpyruvyltransferase
phosphopyruvate-uridine diphosphoacetylglucosaminepyruvatetransferase
pyruvate-UDP-acetylglucosamine transferase
pyruvate-uridine diphospho-N-acetylglucosamine transferase
pyruvate-uridine diphospho-N-acetyl-glucosamine transferase
pyruvic-uridine diphospho-N-acetylglucosaminyltransferase

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
P0A749 MURA_ECOLI Phosphoenolpyruvate + UDP-N-acetyl-D- glucosamine = phosphate + UDP-N-acetyl-3-O-(1-carboxyvinyl)-D- glucosamine. Cytoplasm (Probable).
P33038 MURA_ENTCL Phosphoenolpyruvate + UDP-N-acetyl-D- glucosamine = phosphate + UDP-N-acetyl-3-O-(1-carboxyvinyl)-D- glucosamine. Cytoplasm (Probable).

KEGG Pathways
Map code Pathways E.C.
MAP00530 Aminosugars metabolism

Compound table
Substrates Products Intermediates
KEGG-id C00074 C00043 C00009 C04631
E.C.
Compound Phosphoenolpyruvate UDP-N-acetyl-D-glucosamine Orthophosphate UDP-N-acetyl-3-(1-carboxyvinyl)-D-glucosamine
Type carboxyl group,phosphate group/phosphate ion amide group,carbohydrate,nucleotide phosphate group/phosphate ion amide group,carbohydrate,carboxyl group,nucleotide
ChEBI 44897
 44897
 		16264
 16264
 		26078
 26078
 		68507
 68507
PubChem 1005
 58114173
 59658623
 1005
 58114173
 59658623
 		445675
 445675
 		1004
 22486802
 1004
 22486802
 		172502
 172502
1a2nA01 Unbound Unbound Unbound Unbound Transition-state-analogue:TET
1dlgA01 Unbound Unbound Bound:2xPO4 Unbound Unbound
1dlgB01 Unbound Unbound Bound:2xPO4 Unbound Unbound
1ejcA01 Unbound Unbound Unbound Unbound Unbound
1ejdA01 Unbound Unbound Bound:2xPO4 Unbound Unbound
1ejdB01 Unbound Unbound Bound:5xPO4 Unbound Unbound
1eynA01 Unbound Unbound Unbound Unbound Unbound
1nawA01 Unbound Unbound Unbound Unbound Unbound
1nawB01 Unbound Unbound Unbound Unbound Unbound
1uaeA01 Analogue:FCN Bound:UD1 Unbound Unbound Unbound
1a2nA02 Unbound Unbound Unbound Unbound Unbound
1dlgA02 Unbound Unbound Bound:PO4 Unbound Unbound
1dlgB02 Unbound Unbound Bound:3xPO4 Unbound Unbound
1ejcA02 Unbound Unbound Bound:PO4 Unbound Unbound
1ejdA02 Unbound Unbound Bound:4xPO4 Unbound Unbound
1ejdB02 Unbound Unbound Bound:4xPO4 Unbound Unbound
1eynA02 Unbound Unbound Unbound Unbound Unbound
1nawA02 Unbound Unbound Unbound Unbound Unbound
1nawB02 Unbound Unbound Unbound Unbound Unbound
1uaeA02 Unbound Unbound Unbound Unbound Unbound

Reference for Active-site residues
resource references E.C.
Literature [15] & Swiss-prot;P33038, P0A749

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
1a2nA01 LYS 22;ASN 23; ;ARG 120 mutant C115A
1dlgA01 LYS 22;ASN 23; ;ARG 120 mutant C115S
1dlgB01 LYS 22;ASN 23; ;ARG 120 mutant C115S
1ejcA01 LYS 22;ASN 23;CYS 115;ARG 120
1ejdA01 LYS 22;ASN 23;CYS 115;ARG 120
1ejdB01 LYS 22;ASN 23;CYS 115;ARG 120
1eynA01 LYS 22;ASN 23;CYS 115;ARG 120
1nawA01 LYS 22;ASN 23;CYS 115;ARG 120
1nawB01 LYS 22;ASN 23;CYS 115;ARG 120
1uaeA01 LYS 22;ASN 23;CYS 115;ARG 120
1a2nA02 ASP 305;ARG 397
1dlgA02 ASP 305;ARG 397
1dlgB02 ASP 305;ARG 397
1ejcA02 ASP 305;ARG 397
1ejdA02 ASP 305;ARG 397
1ejdB02 ASP 305;ARG 397
1eynA02 ASP 305;ARG 397
1nawA02 ASP 305;ARG 397
1nawB02 ASP 305;ARG 397
1uaeA02 ASP 305;ARG 397

References for Catalytic Mechanism
References Sections No. of steps in catalysis
[2]
Scheme 1 4
[3]
p.1072-1073
[4]
p.1471
[6]
Scheme 1, Scheme 2, Scheme 3, p.4927-4928 4
[8]
Scheme 3, p.2575-2577 2
[13]
Scheme 1, p.12674-12677 3
[15]
Scheme 1, Scheme 2, p.1556-1558 2
[18]
Fig.3, p.3-4 2

References
[1]
Resource
Comments
Medline ID
PubMed ID 8075064
Journal Biochemistry
Year 1994
Volume 33
Pages 10638-45
Authors Brown ED, Marquardt JL, Lee JP, Walsh CT, Anderson KS
Title Detection and characterization of a phospholactoyl-enzyme adduct in the reaction catalyzed by UDP-N-acetylglucosamine enolpyruvoyl transferase, MurZ.
Related PDB
Related UniProtKB
[2]
Resource
Comments
Medline ID
PubMed ID 7999765
Journal Biochemistry
Year 1994
Volume 33
Pages 15071-9
Authors Ramilo C, Appleyard RJ, Wanke C, Krekel F, Amrhein N, Evans JN
Title Detection of the covalent intermediate of UDP-N-acetylglucosamine enolpyruvyl transferase by solution-state and time-resolved solid-state NMR spectroscopy.
Related PDB
Related UniProtKB
[3]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS)
Medline ID 96398695
PubMed ID 8805592
Journal Structure
Year 1996
Volume 4
Pages 1065-75
Authors Schonbrunn E, Sack S, Eschenburg S, Perrakis A, Krekel F, Amrhein N, Mandelkow E
Title Crystal structure of UDP-N-acetylglucosamine enolpyruvyltransferase, the target of the antibiotic fosfomycin.
Related PDB 1naw
Related UniProtKB P33038
[4]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS)
Medline ID 97148340
PubMed ID 8994972
Journal Structure
Year 1996
Volume 4
Pages 1465-74
Authors Skarzynski T, Mistry A, Wonacott A, Hutchinson SE, Kelly VA, Duncan K
Title Structure of UDP-N-acetylglucosamine enolpyruvyl transferase, an enzyme essential for the synthesis of bacterial peptidoglycan, complexed with substrate UDP-N-acetylglucosamine and the drug fosfomycin.
Related PDB 1uae
Related UniProtKB P0A749
[5]
Resource
Comments
Medline ID
PubMed ID 8776890
Journal J Struct Biol
Year 1996
Volume 117
Pages 73-6
Authors Sack S, Dauter Z, Wanke C, Amrhein N, Mandelkow E, Schonbrunn E
Title Crystallization and preliminary X-ray diffraction analysis of UDP-N-acetylglucosamine enolpyruvyltransferase of Enterobacter cloacae.
Related PDB
Related UniProtKB
[6]
Resource
Comments
Medline ID
PubMed ID 8664284
Journal Biochemistry
Year 1996
Volume 35
Pages 4923-8
Authors Kim DH, Lees WJ, Kempsell KE, Lane WS, Duncan K, Walsh CT
Title 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.
Related PDB
Related UniProtKB
[7]
Resource
Comments
Medline ID
PubMed ID 9654090
Journal Eur J Biochem
Year 1998
Volume 253
Pages 406-12
Authors Schonbrunn E, Svergun DI, Amrhein N, Koch MH
Title Studies on the conformational changes in the bacterial cell wall biosynthetic enzyme UDP-N-acetylglucosamine enolpyruvyltransferase (MurA).
Related PDB
Related UniProtKB
[8]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.8 ANGSTROMS)
Medline ID 98153140
PubMed ID 9485407
Journal Biochemistry
Year 1998
Volume 37
Pages 2572-7
Authors Skarzynski T, Kim DH, Lees WJ, Walsh CT, Duncan K
Title Stereochemical course of enzymatic enolpyruvyl transfer and catalytic conformation of the active site revealed by the crystal structure of the fluorinated analogue of the reaction tetrahedral intermediate bound to the active site of the C115A mutant of MurA.
Related PDB 1a2n
Related UniProtKB P0A749
[9]
Resource
Comments
Medline ID
PubMed ID 10413459
Journal Biochemistry
Year 1999
Volume 38
Pages 8864-78
Authors Krekel F, Oecking C, Amrhein N, Macheroux P
Title Substrate and inhibitor-induced conformational changes in the structurally related enzymes UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) and 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS).
Related PDB
Related UniProtKB
[10]
Resource
Comments
Medline ID
PubMed ID 10529188
Journal Biochemistry
Year 1999
Volume 38
Pages 13162-9
Authors Samland AK, Amrhein N, Macheroux P
Title Lysine 22 in UDP-N-acetylglucosamine enolpyruvyl transferase from Enterobacter cloacae is crucial for enzymatic activity and the formation of covalent adducts with the substrate phosphoenolpyruvate and the antibiotic fosfomycin.
Related PDB
Related UniProtKB
[11]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 10842342
Journal Proteins
Year 2000
Volume 40
Pages 290-8
Authors Eschenburg S, Schonbrunn E
Title Comparative X-ray analysis of the un-liganded fosfomycin-target murA.
Related PDB 1ejc 1ejd
Related UniProtKB
[12]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 10823915
Journal Proc Natl Acad Sci U S A
Year 2000
Volume 97
Pages 6345-9
Authors Schonbrunn E, Eschenburg S, Luger K, Kabsch W, Amrhein N
Title Structural basis for the interaction of the fluorescence probe 8-anilino-1-naphthalene sulfonate (ANS) with the antibiotic target MurA.
Related PDB 1eyn
Related UniProtKB
[13]
Resource
Comments
Medline ID
PubMed ID 11027147
Journal Biochemistry
Year 2000
Volume 39
Pages 12671-7
Authors Krekel F, Samland AK, Macheroux P, Amrhein N, Evans JN
Title Determination of the pKa value of C115 in MurA (UDP-N-acetylglucosamine enolpyruvyltransferase) from Enterobacter cloacae.
Related PDB
Related UniProtKB
[14]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS)
Medline ID 20160492
PubMed ID 10694381
Journal Biochemistry
Year 2000
Volume 39
Pages 2164-73
Authors Schonbrunn E, Eschenburg S, Krekel F, Luger K, Amrhein N
Title Role of the loop containing residue 115 in the induced-fit mechanism of the bacterial cell wall biosynthetic enzyme MurA.
Related PDB 1dlg
Related UniProtKB P33038
[15]
Resource
Comments
Medline ID
PubMed ID 11327813
Journal Biochemistry
Year 2001
Volume 40
Pages 1550-9
Authors Samland AK, Etezady-Esfarjani T, Amrhein N, Macheroux P
Title Asparagine 23 and aspartate 305 are essential residues in the active site of UDP-N-acetylglucosamine enolpyruvyl transferase from Enterobacter cloacae.
Related PDB
Related UniProtKB
[16]
Resource
Comments
Medline ID
PubMed ID 11502190
Journal Biochemistry
Year 2001
Volume 40
Pages 9950-6
Authors Samland AK, Jelesarov I, Kuhn R, Amrhein N, Macheroux P
Title Thermodynamic characterization of ligand-induced conformational changes in UDP-N-acetylglucosamine enolpyruvyl transferase.
Related PDB
Related UniProtKB
[17]
Resource
Comments
Medline ID
PubMed ID 11600375
Journal Antimicrob Agents Chemother
Year 2001
Volume 45
Pages 3182-8
Authors Baum EZ, Montenegro DA, Licata L, Turchi I, Webb GC, Foleno BD, Bush K
Title Identification and characterization of new inhibitors of the Escherichia coli MurA enzyme.
Related PDB
Related UniProtKB
[18]
Resource
Comments
Medline ID
PubMed ID 12492849
Journal Mol Microbiol
Year 2003
Volume 47
Pages 1-12
Authors El Zoeiby A, Sanschagrin F, Levesque RC
Title Structure and function of the Mur enzymes: development of novel inhibitors.
Related PDB
Related UniProtKB

Comments
According to the literature [8] & [15], this enzyme catalyzes two successive reactions, Addition and Elimination, as follows:
(A) Additive double-bond deformation (Addition of hydroxyl group to sp2 Carbon):
(A1) A general base (Asp305) activates the acceptor, 3'-hydroxyl group of UDP-NAG, by deprotonating the group.
(A2) The activated acceptor group would then make a nucleophilic attack on the C-2 atom of phosphoenolpyruvate (PEP).
(A3) Simultanesouly, Cys115 would act as a general acid, to protonate the C-3 atom of PEP, of which double bond would change into single bond, resulting in the formation of the tetrahedral intermediate. This catalysis proceeds by SN2-reaction.
(A4) Here, the interemediate can be stabilized by Asn23, Lys22, Arg120 and Arg397, as well as by Asp305 (see [8] & [15]). In particular, the added hydroxyl oxygen seems to be stabilized by Asn23.
(B) Eliminative double-bond formation (Elimination of phosphate group):
(B1) Cys115 would act as a general base to abstract a proton from the C-3 atom of the transferred group, recovering the double bond and facilitating the dissociation of the leaving phosphate group.
(B2) The leaving phosphate seems to be stabilized by the positive charges of Lys22, Arg120 and Arg397.
However, the function of cysteine residue as a general acid is quite unusual, and some papers such as [6] reported that Cys115 could act as a nucleophile, forming a covalent intermediate (O-phosphothioketal intermediate). The literature [13] determined the pKa value of Cys115, and suggested that the residue can be a proton donor in the catalysis, but mentioned that the possibility of its role as nucleophile could still not be ruled out. On the other hand, other papers, [6] & [18], suggested that the formation of the covalent intermediate would not be required for the main catalytic pathway.

Created Updated
2002-11-25 2009-02-26