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Escherichia coli K-12 substr. MG1655 Enzyme: UDP-N-acetylglucosamine enolpyruvoyl transferase



Gene: murA Accession Numbers: EG11358 (EcoCyc), b3189, ECK3178

Synonyms: mrbA, murZ

Regulation Summary Diagram: ?

Summary:
UDP-N-acetylglucosamine enolpyruvoyl transferase (MurA) catalyzes the first committed step in the assembly of peptidoglycan [Marquardt92]. In exponentially growing cells, the intracellular concentration of UDP-N-acetylglucosamine is approximately 100 µM, while the concentration of the MurA product, UDP-N-acetylglucosamine enolpyruvate is only 2 µM [MenginLecreulx83]. MurA activity is feedback-inhibited by UDP-MurNAc, the product of the second enzyme in the UDP-N-acetylmuramoyl-pentapeptide biosynthesis I (meso-DAP-containing) pathway [Mizyed05].

The reaction mechanism of the enzyme has been investigated. A tetrahedral adduct can be formed at the active site of the enzyme during catalyis [Brown94a, Marquardt93, Kim95e]. However, covalent enzyme adduct formation is not required for catalysis [Kim96j]. The stereochemical course of the reaction has been investigated [Skarzynski98, Lees95, Skarzynski98]. Catalysis involves conformational transitions within the enzyme [Jackson09].

MurA is the site of action for the antibiotic fosfomycin. Inactivation of MurA is dependent on the presence of the substrate UDP-GlcNAc [Marquardt94]. The antibiotic covalently binds to the active site nucleophile Cys115 of the enzyme [Marquardt94, Skarzynski96]. A C115D mutant is active, but resistant to fosfomycin [Kim96j]. Overexpression of MurA increases resistance to fosfomycin [Xu06e, Couce12]; it is the only chromosomally encoded protein whose overexpression yields fosfomycin resistance [Couce12]. A murA mutant with increased resistance to fosfomycin was identified [Venkateswaran72]; the temperature-sensitive phenotype of that mutant may be unrelated to the murA mutation [Brown95b]. Novel inhibitors of MurA activity have been sought and investigated [Baum01, Barbosa02b, Francisco04, Bachelier06, Dunsmore08, Miller10]. MurA is also the primary physiological target of the nonnatural toxin bromoacetate [Desai10].

Crystal structures of wild-type and mutant MurA in various complexes have been solved [Skarzynski96, Skarzynski98, Steinbach08, Jackson09, Han10d]. Reinterpretation of existing crystal structure data suggests that under physiological conditions, MurA exists in a dormant complex containing UDP-N-acetylmuramic acid (UNAM) and the covalent PEP adduct at Cys115 [Zhu12a].

murA is an essential gene [Brown95b, Goh09]. A strain containing a conditionally lethal amber mutation in murA has been generated [Herring04a].

Reviews: [El03, Barreteau08, Typas12]

Citations: [Nicolaou12]

Gene Citations: [Guinote12]

Locations: cytosol

Map Position: [3,333,257 <- 3,334,516] (71.84 centisomes)
Length: 1260 bp / 419 aa

Molecular Weight of Polypeptide: 44.818 kD (from nucleotide sequence), 44.0 kD (experimental) [Marquardt92 ]

pI: 6.13

Unification Links: ASAP:ABE-0010479 , CGSC:33518 , DIP:DIP-48060N , EchoBASE:EB1333 , EcoGene:EG11358 , EcoliWiki:b3189 , Mint:MINT-1224932 , ModBase:P0A749 , OU-Microarray:b3189 , PortEco:murA , PR:PRO_000023312 , Pride:P0A749 , Protein Model Portal:P0A749 , RefSeq:NP_417656 , RegulonDB:EG11358 , SMR:P0A749 , String:511145.b3189 , UniProt:P0A749

Relationship Links: InterPro:IN-FAMILY:IPR001986 , InterPro:IN-FAMILY:IPR005750 , InterPro:IN-FAMILY:IPR013792 , Panther:IN-FAMILY:PTHR21090:SF4 , PDB:Structure:1A2N , PDB:Structure:1UAE , PDB:Structure:2Z2C , PDB:Structure:3ISS , PDB:Structure:3KQJ , PDB:Structure:3KR6 , PDB:Structure:3SWD , Pfam:IN-FAMILY:PF00275

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0009252 - peptidoglycan biosynthetic process Inferred from experiment Inferred by computational analysis [UniProtGOA12, UniProtGOA11, GOA06, Venkateswaran72]
GO:0007049 - cell cycle Inferred by computational analysis [UniProtGOA11]
GO:0008360 - regulation of cell shape Inferred by computational analysis [UniProtGOA11]
GO:0019277 - UDP-N-acetylgalactosamine biosynthetic process Inferred by computational analysis [GOA01]
GO:0051301 - cell division Inferred by computational analysis [UniProtGOA11]
GO:0071555 - cell wall organization Inferred by computational analysis [UniProtGOA11]
Molecular Function: GO:0008760 - UDP-N-acetylglucosamine 1-carboxyvinyltransferase activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01a, GOA01, Marquardt92, Brown94a]
GO:0003824 - catalytic activity Inferred by computational analysis [GOA01]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11]
GO:0016765 - transferase activity, transferring alkyl or aryl (other than methyl) groups Inferred by computational analysis [GOA01]
Cellular Component: GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [DiazMejia09, Ishihama08, LopezCampistrou05]
GO:0005737 - cytoplasm Inferred by computational analysis [UniProtGOA11a, UniProtGOA11, GOA06]

MultiFun Terms: cell structure murein
metabolism biosynthesis of macromolecules (cellular constituents) murein (peptidoglycan)

Essentiality data for murA knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox No 37 Aerobic 7   No [Baba06, Comment 2]

Credits:
Last-Curated ? 15-Jul-2013 by Keseler I , SRI International


Enzymatic reaction of: UDP-N-acetylglucosamine enolpyruvoyl transferase

Synonyms: enoylpyruvate transferase, UDP-N-acetylglucosamine 1-carboxyvinyltransferase, phosphoenolpyruvate:UDP-N-acetyl-D-glucosamine 1-carboxyvinyltransferase, enolpyruvyl UDP-GlcNAc synthase

EC Number: 2.5.1.7

UDP-N-acetyl-α-D-glucosamine + phosphoenolpyruvate <=> UDP-N-acetyl-α-D-glucosamine-enolpyruvate + phosphate

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the Enzyme Commission system.

The reaction is favored in the direction shown.

In Pathways: peptidoglycan biosynthesis I (meso-diaminopimelate containing) , UDP-N-acetylmuramoyl-pentapeptide biosynthesis I (meso-DAP-containing)

Inhibitors (Competitive): UDP-N-α-D-acetylmuramate [Mizyed05, Zhu12a]

Inhibitors (Irreversible): bromoacetate [Desai10] , fosfomycin [Venkateswaran72, Marquardt94]

Inhibitors (Unknown Mechanism): UNAG-3P [Mutschler11]

Primary Physiological Regulators of Enzyme Activity: UDP-N-α-D-acetylmuramate

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
phosphoenolpyruvate
4.0
[Gautam11, BRENDA14]
phosphoenolpyruvate
1000.0
[Marquardt92, BRENDA14]
phosphoenolpyruvate
4.1
[Dai02, BRENDA14]
phosphoenolpyruvate
0.4, 0.4
[Kim96j, BRENDA14]
UDP-N-acetyl-α-D-glucosamine
15.0
[Brown94a, BRENDA14]
UDP-N-acetyl-α-D-glucosamine
15.0
3.8
[Gautam11, BRENDA14]
UDP-N-acetyl-α-D-glucosamine
4.75
[Marquardt92, BRENDA14]

pH(opt): 5.5-9 [Kim96j]


Sequence Features

Feature Class Location Citations Comment
Modified-Residue 115
[UniProt10]
UniProt: 2-(S-cysteinyl)pyruvic acid O- phosphothioketal;
Active-Site 115
[UniProt10]
UniProt: Proton donor;


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
10/20/97 Gene b3189 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG11358; confirmed by SwissProt match.


References

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

Bachelier06: Bachelier A, Mayer R, Klein CD (2006). "Sesquiterpene lactones are potent and irreversible inhibitors of the antibacterial target enzyme MurA." Bioorg Med Chem Lett 16(21);5605-9. PMID: 16945528

Barbosa02b: Barbosa MD, Yang G, Fang J, Kurilla MG, Pompliano DL (2002). "Development of a whole-cell assay for peptidoglycan biosynthesis inhibitors." Antimicrob Agents Chemother 46(4);943-6. PMID: 11897573

Barreteau08: Barreteau H, Kovac A, Boniface A, Sova M, Gobec S, Blanot D (2008). "Cytoplasmic steps of peptidoglycan biosynthesis." FEMS Microbiol Rev 32(2);168-207. PMID: 18266853

Baum01: Baum EZ, Montenegro DA, Licata L, Turchi I, Webb GC, Foleno BD, Bush K (2001). "Identification and characterization of new inhibitors of the Escherichia coli MurA enzyme." Antimicrob Agents Chemother 45(11);3182-8. PMID: 11600375

BRENDA14: BRENDA team (2014). "Imported from BRENDA version existing on Aug 2014." http://www.brenda-enzymes.org.

Brown94a: Brown ED, Marquardt JL, Lee JP, Walsh CT, Anderson KS (1994). "Detection and characterization of a phospholactoyl-enzyme adduct in the reaction catalyzed by UDP-N-acetylglucosamine enolpyruvoyl transferase, MurZ." Biochemistry 1994;33(35);10638-45. PMID: 8075064

Brown95b: Brown ED, Vivas EI, Walsh CT, Kolter R (1995). "MurA (MurZ), the enzyme that catalyzes the first committed step in peptidoglycan biosynthesis, is essential in Escherichia coli." J Bacteriol 177(14);4194-7. PMID: 7608103

Couce12: Couce A, Briales A, Rodriguez-Rojas A, Costas C, Pascual A, Blazquez J (2012). "Genomewide overexpression screen for fosfomycin resistance in Escherichia coli: MurA confers clinical resistance at low fitness cost." Antimicrob Agents Chemother 56(5);2767-9. PMID: 22371901

Dai02: Dai HJ, Parker CN, Bao JJ (2002). "Characterization and inhibition study of MurA enzyme by capillary electrophoresis." J Chromatogr B Analyt Technol Biomed Life Sci 766(1);123-32. PMID: 11820287

Desai10: Desai KK, Miller BG (2010). "Recruitment of genes and enzymes conferring resistance to the nonnatural toxin bromoacetate." Proc Natl Acad Sci U S A 107(42);17968-73. PMID: 20921376

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

Dunsmore08: Dunsmore CJ, Miller K, Blake KL, Patching SG, Henderson PJ, Garnett JA, Stubbings WJ, Phillips SE, Palestrant DJ, Angeles Jde L, Leeds JA, Chopra I, Fishwick CW (2008). "2-Aminotetralones: novel inhibitors of MurA and MurZ." Bioorg Med Chem Lett 18(5);1730-4. PMID: 18261901

El03: El Zoeiby A, Sanschagrin F, Levesque RC (2003). "Structure and function of the Mur enzymes: development of novel inhibitors." Mol Microbiol 47(1);1-12. PMID: 12492849

Francisco04: Francisco GD, Li Z, Albright JD, Eudy NH, Katz AH, Petersen PJ, Labthavikul P, Singh G, Yang Y, Rasmussen BA, Lin YI, Mansour TS (2004). "Phenyl thiazolyl urea and carbamate derivatives as new inhibitors of bacterial cell-wall biosynthesis." Bioorg Med Chem Lett 14(1);235-8. PMID: 14684334

Gautam11: Gautam A, Rishi P, Tewari R (2011). "UDP-N-acetylglucosamine enolpyruvyl transferase as a potential target for antibacterial chemotherapy: recent developments." Appl Microbiol Biotechnol 92(2);211-25. PMID: 21822642

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

GOA01: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA01a: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

Goh09: Goh S, Boberek JM, Nakashima N, Stach J, Good L (2009). "Concurrent growth rate and transcript analyses reveal essential gene stringency in Escherichia coli." PLoS One 4(6);e6061. PMID: 19557168

Guinote12: Guinote IB, Moreira RN, Freire P, Arraiano CM (2012). "Characterization of the BolA Homolog IbaG: A New Gene Involved in Acid Resistance." J Microbiol Biotechnol 22(4);484-93. PMID: 22534295

Han10d: Han H, Yang Y, Olesen SH, Becker A, Betzi S, Schonbrunn E (2010). "The fungal product terreic acid is a covalent inhibitor of the bacterial cell wall biosynthetic enzyme UDP-N-acetylglucosamine 1-carboxyvinyltransferase (MurA)." Biochemistry 49(19);4276-82. PMID: 20392080

Herring04a: Herring CD, Blattner FR (2004). "Conditional lethal amber mutations in essential Escherichia coli genes." J Bacteriol 186(9);2673-81. PMID: 15090508

Ishihama08: Ishihama Y, Schmidt T, Rappsilber J, Mann M, Hartl FU, Kerner MJ, Frishman D (2008). "Protein abundance profiling of the Escherichia coli cytosol." BMC Genomics 9;102. PMID: 18304323

Jackson09: Jackson SG, Zhang F, Chindemi P, Junop MS, Berti PJ (2009). "Evidence of kinetic control of ligand binding and staged product release in MurA (enolpyruvyl UDP-GlcNAc synthase)-catalyzed reactions." Biochemistry 48(49);11715-23. PMID: 19899805

Kim95e: Kim DH, Lees WJ, Walsh CT (1995). "Stereochemical analysis of the tetrahedral adduct formed at the active site of UDP-GlcNAc enolpyruvyl transferase from the pseudosubstrates, (E)- and (Z)-3-fluorophosphoenolpyruvate, in D2O." J Am Chem Soc 117:6380-6381.

Kim96j: Kim DH, Lees WJ, Kempsell KE, Lane WS, Duncan K, Walsh CT (1996). "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." Biochemistry 35(15);4923-8. PMID: 8664284

Lees95: Lees WJ, Walsh CT (1995). "Analysis of the enol ether transfer catalyzed by UDP-GlcNAc enolpyruvyl transferase using (E)- and (Z)-isomers of phosphoenolbutyrate: stereochemical, partitioning, and isotope effect studies." J Am Chem Soc 117:7329-7337.

LopezCampistrou05: Lopez-Campistrous A, Semchuk P, Burke L, Palmer-Stone T, Brokx SJ, Broderick G, Bottorff D, Bolch S, Weiner JH, Ellison MJ (2005). "Localization, annotation, and comparison of the Escherichia coli K-12 proteome under two states of growth." Mol Cell Proteomics 4(8);1205-9. PMID: 15911532

Marquardt92: Marquardt JL, Siegele DA, Kolter R, Walsh CT (1992). "Cloning and sequencing of Escherichia coli murZ and purification of its product, a UDP-N-acetylglucosamine enolpyruvyl transferase." J Bacteriol 1992;174(17);5748-52. PMID: 1512209

Marquardt93: Marquardt JL, Brown ED, Walsh CT, Anderson KS (1993). "Isolation and structural elucidation of a tetrahedral intermediate in the UDP-N-acetylglucosamine enolpyruvoyl transferase enzymatic pathway." J Am Chem Soc 115:10398-10399.

Marquardt94: Marquardt JL, Brown ED, Lane WS, Haley TM, Ichikawa Y, Wong CH, Walsh CT (1994). "Kinetics, stoichiometry, and identification of the reactive thiolate in the inactivation of UDP-GlcNAc enolpyruvoyl transferase by the antibiotic fosfomycin." Biochemistry 1994;33(35);10646-51. PMID: 8075065

MenginLecreulx83: Mengin-Lecreulx D, Flouret B, van Heijenoort J (1983). "Pool levels of UDP N-acetylglucosamine and UDP N-acetylglucosamine-enolpyruvate in Escherichia coli and correlation with peptidoglycan synthesis." J Bacteriol 154(3);1284-90. PMID: 6222035

Miller10: Miller K, Dunsmore CJ, Leeds JA, Patching SG, Sachdeva M, Blake KL, Stubbings WJ, Simmons KJ, Henderson PJ, De Los Angeles J, Fishwick CW, Chopra I (2010). "Benzothioxalone derivatives as novel inhibitors of UDP-N-acetylglucosamine enolpyruvyl transferases (MurA and MurZ)." J Antimicrob Chemother 65(12);2566-73. PMID: 20861142

Mizyed05: Mizyed S, Oddone A, Byczynski B, Hughes DW, Berti PJ (2005). "UDP-N-acetylmuramic acid (UDP-MurNAc) is a potent inhibitor of MurA (enolpyruvyl-UDP-GlcNAc synthase)." Biochemistry 44(10);4011-7. PMID: 15751977

Mutschler11: Mutschler H, Gebhardt M, Shoeman RL, Meinhart A (2011). "A novel mechanism of programmed cell death in bacteria by toxin-antitoxin systems corrupts peptidoglycan synthesis." PLoS Biol 9(3);e1001033. PMID: 21445328

Nicolaou12: Nicolaou SA, Gaida SM, Papoutsakis ET (2012). "Exploring the combinatorial genomic space in Escherichia coli for ethanol tolerance." Biotechnol J 7(11);1337-45. PMID: 22847918

Skarzynski96: Skarzynski T, Mistry A, Wonacott A, Hutchinson SE, Kelly VA, Duncan K (1996). "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." Structure 4(12);1465-74. PMID: 8994972

Skarzynski98: Skarzynski T, Kim DH, Lees WJ, Walsh CT, Duncan K (1998). "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." Biochemistry 37(8);2572-7. PMID: 9485407

Steinbach08: Steinbach A, Scheidig AJ, Klein CD (2008). "The unusual binding mode of cnicin to the antibacterial target enzyme MurA revealed by X-ray crystallography." J Med Chem 51(16);5143-7. PMID: 18672863

Typas12: Typas A, Banzhaf M, Gross CA, Vollmer W (2012). "From the regulation of peptidoglycan synthesis to bacterial growth and morphology." Nat Rev Microbiol 10(2);123-36. PMID: 22203377

UniProt10: UniProt Consortium (2010). "UniProt version 2010-11 released on 2010-11-02 00:00:00." Database.

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."

UniProtGOA12: UniProt-GOA (2012). "Gene Ontology annotation based on UniPathway vocabulary mapping."

Venkateswaran72: Venkateswaran PS, Wu HC (1972). "Isolation and characterization of a phosphonomycin-resistant mutant of Escherichia coli K-12." J Bacteriol 110(3);935-44. PMID: 4555418

Xu06e: Xu HH, Real L, Bailey MW (2006). "An array of Escherichia coli clones over-expressing essential proteins: a new strategy of identifying cellular targets of potent antibacterial compounds." Biochem Biophys Res Commun 349(4);1250-7. PMID: 16978582

Zhu12a: Zhu JY, Yang Y, Han H, Betzi S, Olesen SH, Marsilio F, Schonbrunn E (2012). "Functional consequence of covalent reaction of phosphoenolpyruvate with UDP-N-acetylglucosamine 1-carboxyvinyltransferase (MurA)." J Biol Chem 287(16);12657-67. PMID: 22378791


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Tue Nov 25, 2014, BIOCYC13A.