EzCatDB: D00415

DB codeD00415
RLCP classification4.23.18400.69
5.303.667700.54
CATH domainDomain 13.65.10.10Catalytic domain
Domain 23.65.10.10Catalytic domain
E.C.2.5.1.7
CSA1uae


Enzyme Name
Swiss-protKEGG

P0A749P33038
Protein nameUDP-N-acetylglucosamine 1-carboxyvinyltransferaseUDP-N-acetylglucosamine 1-carboxyvinyltransferaseUDP-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
SynonymsEC 2.5.1.7
Enoylpyruvate transferase
UDP-N-acetylglucosamine enolpyruvyl transferase
EPT
EC 2.5.1.7
Enoylpyruvate transferase
UDP-N-acetylglucosamine enolpyruvyl transferase
EPT

KEGG pathways
MAP codePathways
MAP00530Aminosugars metabolism

Swiss-prot:Accession NumberP0A749P33038
Entry nameMURA_ECOLIMURA_ENTCL
ActivityPhosphoenolpyruvate + UDP-N-acetyl-D- glucosamine = phosphate + UDP-N-acetyl-3-O-(1-carboxyvinyl)-D- glucosamine.Phosphoenolpyruvate + UDP-N-acetyl-D- glucosamine = phosphate + UDP-N-acetyl-3-O-(1-carboxyvinyl)-D- glucosamine.
Subunit

Subcellular locationCytoplasm (Probable).Cytoplasm (Probable).
Cofactor



SubstratesProductsintermediates
KEGG-idC00074C00043C00009C04631
CompoundPhosphoenolpyruvateUDP-N-acetyl-D-glucosamineOrthophosphateUDP-N-acetyl-3-(1-carboxyvinyl)-D-glucosamine
Typecarboxyl group,phosphate group/phosphate ionamide group,carbohydrate,nucleotidephosphate group/phosphate ionamide group,carbohydrate,carboxyl group,nucleotide
1a2nA01UnboundUnboundUnboundUnboundTransition-state-analogue:TET
1dlgA01UnboundUnboundBound:2xPO4UnboundUnbound
1dlgB01UnboundUnboundBound:2xPO4UnboundUnbound
1ejcA01UnboundUnboundUnboundUnboundUnbound
1ejdA01UnboundUnboundBound:2xPO4UnboundUnbound
1ejdB01UnboundUnboundBound:5xPO4UnboundUnbound
1eynA01UnboundUnboundUnboundUnboundUnbound
1nawA01UnboundUnboundUnboundUnboundUnbound
1nawB01UnboundUnboundUnboundUnboundUnbound
1uaeA01Analogue:FCNBound:UD1UnboundUnboundUnbound
1a2nA02UnboundUnboundUnboundUnboundUnbound
1dlgA02UnboundUnboundBound:PO4UnboundUnbound
1dlgB02UnboundUnboundBound:3xPO4UnboundUnbound
1ejcA02UnboundUnboundBound:PO4UnboundUnbound
1ejdA02UnboundUnboundBound:4xPO4UnboundUnbound
1ejdB02UnboundUnboundBound:4xPO4UnboundUnbound
1eynA02UnboundUnboundUnboundUnboundUnbound
1nawA02UnboundUnboundUnboundUnboundUnbound
1nawB02UnboundUnboundUnboundUnboundUnbound
1uaeA02UnboundUnboundUnboundUnboundUnbound

Active-site residues
resource
Literature [15] & Swiss-prot;P33038, P0A749
pdbCatalytic residuescomment
1a2nA01LYS 22;ASN 23;       ;ARG 120
mutant C115A
1dlgA01LYS 22;ASN 23;       ;ARG 120
mutant C115S
1dlgB01LYS 22;ASN 23;       ;ARG 120
mutant C115S
1ejcA01LYS 22;ASN 23;CYS 115;ARG 120

1ejdA01LYS 22;ASN 23;CYS 115;ARG 120

1ejdB01LYS 22;ASN 23;CYS 115;ARG 120

1eynA01LYS 22;ASN 23;CYS 115;ARG 120

1nawA01LYS 22;ASN 23;CYS 115;ARG 120

1nawB01LYS 22;ASN 23;CYS 115;ARG 120

1uaeA01LYS 22;ASN 23;CYS 115;ARG 120

1a2nA02ASP 305;ARG 397

1dlgA02ASP 305;ARG 397

1dlgB02ASP 305;ARG 397

1ejcA02ASP 305;ARG 397

1ejdA02ASP 305;ARG 397

1ejdB02ASP 305;ARG 397

1eynA02ASP 305;ARG 397

1nawA02ASP 305;ARG 397

1nawB02ASP 305;ARG 397

1uaeA02ASP 305;ARG 397


References for Catalytic Mechanism
ReferencesSectionsNo. of steps in catalysis
[2]Scheme 14
[3]p.1072-1073
[4]p.1471
[6]Scheme 1, Scheme 2, Scheme 3, p.4927-49284
[8]Scheme 3, p.2575-25772
[13]Scheme 1, p.12674-126773
[15]Scheme 1, Scheme 2, p.1556-15582
[18]Fig.3, p.3-42

references
[1]
PubMed ID8075064
JournalBiochemistry
Year1994
Volume33
Pages10638-45
AuthorsBrown ED, Marquardt JL, Lee JP, Walsh CT, Anderson KS
TitleDetection and characterization of a phospholactoyl-enzyme adduct in the reaction catalyzed by UDP-N-acetylglucosamine enolpyruvoyl transferase, MurZ.
[2]
PubMed ID7999765
JournalBiochemistry
Year1994
Volume33
Pages15071-9
AuthorsRamilo C, Appleyard RJ, Wanke C, Krekel F, Amrhein N, Evans JN
TitleDetection of the covalent intermediate of UDP-N-acetylglucosamine enolpyruvyl transferase by solution-state and time-resolved solid-state NMR spectroscopy.
[3]
CommentsX-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS)
Medline ID96398695
PubMed ID8805592
JournalStructure
Year1996
Volume4
Pages1065-75
AuthorsSchonbrunn E, Sack S, Eschenburg S, Perrakis A, Krekel F, Amrhein N, Mandelkow E
TitleCrystal structure of UDP-N-acetylglucosamine enolpyruvyltransferase, the target of the antibiotic fosfomycin.
Related PDB1naw
Related Swiss-protP33038
[4]
CommentsX-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS)
Medline ID97148340
PubMed ID8994972
JournalStructure
Year1996
Volume4
Pages1465-74
AuthorsSkarzynski T, Mistry A, Wonacott A, Hutchinson SE, Kelly VA, Duncan K
TitleStructure 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 PDB1uae
Related Swiss-protP0A749
[5]
PubMed ID8776890
JournalJ Struct Biol
Year1996
Volume117
Pages73-6
AuthorsSack S, Dauter Z, Wanke C, Amrhein N, Mandelkow E, Schonbrunn E
TitleCrystallization and preliminary X-ray diffraction analysis of UDP-N-acetylglucosamine enolpyruvyltransferase of Enterobacter cloacae.
[6]
PubMed ID8664284
JournalBiochemistry
Year1996
Volume35
Pages4923-8
AuthorsKim DH, Lees WJ, Kempsell KE, Lane WS, Duncan K, Walsh CT
TitleCharacterization 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.
[7]
PubMed ID9654090
JournalEur J Biochem
Year1998
Volume253
Pages406-12
AuthorsSchonbrunn E, Svergun DI, Amrhein N, Koch MH
TitleStudies on the conformational changes in the bacterial cell wall biosynthetic enzyme UDP-N-acetylglucosamine enolpyruvyltransferase (MurA).
[8]
CommentsX-RAY CRYSTALLOGRAPHY (2.8 ANGSTROMS)
Medline ID98153140
PubMed ID9485407
JournalBiochemistry
Year1998
Volume37
Pages2572-7
AuthorsSkarzynski T, Kim DH, Lees WJ, Walsh CT, Duncan K
TitleStereochemical 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 PDB1a2n
Related Swiss-protP0A749
[9]
PubMed ID10413459
JournalBiochemistry
Year1999
Volume38
Pages8864-78
AuthorsKrekel F, Oecking C, Amrhein N, Macheroux P
TitleSubstrate and inhibitor-induced conformational changes in the structurally related enzymes UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) and 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS).
[10]
PubMed ID10529188
JournalBiochemistry
Year1999
Volume38
Pages13162-9
AuthorsSamland AK, Amrhein N, Macheroux P
TitleLysine 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.
[11]
CommentsX-ray crystallography
PubMed ID10842342
JournalProteins
Year2000
Volume40
Pages290-8
AuthorsEschenburg S, Schonbrunn E
TitleComparative X-ray analysis of the un-liganded fosfomycin-target murA.
Related PDB1ejc,1ejd
[12]
CommentsX-ray crystallography
PubMed ID10823915
JournalProc Natl Acad Sci U S A
Year2000
Volume97
Pages6345-9
AuthorsSchonbrunn E, Eschenburg S, Luger K, Kabsch W, Amrhein N
TitleStructural basis for the interaction of the fluorescence probe 8-anilino-1-naphthalene sulfonate (ANS) with the antibiotic target MurA.
Related PDB1eyn
[13]
PubMed ID11027147
JournalBiochemistry
Year2000
Volume39
Pages12671-7
AuthorsKrekel F, Samland AK, Macheroux P, Amrhein N, Evans JN
TitleDetermination of the pKa value of C115 in MurA (UDP-N-acetylglucosamine enolpyruvyltransferase) from Enterobacter cloacae.
[14]
CommentsX-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS)
Medline ID20160492
PubMed ID10694381
JournalBiochemistry
Year2000
Volume39
Pages2164-73
AuthorsSchonbrunn E, Eschenburg S, Krekel F, Luger K, Amrhein N
TitleRole of the loop containing residue 115 in the induced-fit mechanism of the bacterial cell wall biosynthetic enzyme MurA.
Related PDB1dlg
Related Swiss-protP33038
[15]
PubMed ID11327813
JournalBiochemistry
Year2001
Volume40
Pages1550-9
AuthorsSamland AK, Etezady-Esfarjani T, Amrhein N, Macheroux P
TitleAsparagine 23 and aspartate 305 are essential residues in the active site of UDP-N-acetylglucosamine enolpyruvyl transferase from Enterobacter cloacae.
[16]
PubMed ID11502190
JournalBiochemistry
Year2001
Volume40
Pages9950-6
AuthorsSamland AK, Jelesarov I, Kuhn R, Amrhein N, Macheroux P
TitleThermodynamic characterization of ligand-induced conformational changes in UDP-N-acetylglucosamine enolpyruvyl transferase.
[17]
PubMed ID11600375
JournalAntimicrob Agents Chemother
Year2001
Volume45
Pages3182-8
AuthorsBaum EZ, Montenegro DA, Licata L, Turchi I, Webb GC, Foleno BD, Bush K
TitleIdentification and characterization of new inhibitors of the Escherichia coli MurA enzyme.
[18]
PubMed ID12492849
JournalMol Microbiol
Year2003
Volume47
Pages1-12
AuthorsEl Zoeiby A, Sanschagrin F, Levesque RC
TitleStructure and function of the Mur enzymes: development of novel inhibitors.

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.

createdupdated
2002-11-252009-02-26
Copyright: Nozomi Nagano, JST & CBRC-AIST
Funded by PRESTO/Japan Science and Technology Corporation (JST) (December 2001 - November 2004)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2006)
Funded by Grant-in-Aid for Scientific Research (B)/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2008)
Funded by BIRD/Japan Science and Technology Corporation (JST) (September 2005 - September 2010)
Funded by BIRD/Japan Science and Technology Corporation (JST) (October 2007 - September 2010)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2011 - March 2012)

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