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Escherichia coli K-12 substr. MG1655 Enzyme: UDP-N-acetylmuramate-alanine ligase



Gene: murC Accession Numbers: EG10619 (EcoCyc), b0091, ECK0092

Regulation Summary Diagram: ?

Subunit composition of UDP-N-acetylmuramate-alanine ligase = [MurC]2
         UDP-N-acetylmuramate-alanine ligase = MurC

Summary:
UDP-N-acetylmuramate-alanine ligase (MurC) is responsible for the addition of the first amino acid of the peptide moiety in the assembly of the monomer unit of peptidoglycan [Liger95].

The formation of the peptide bond involves a covalent acyl-phosphate intermediate between the carboxyl group of UDP-N-acetylmuramate and the γ phosphate of ATP [Falk96, Bouhss99]. Kinetic data are consistent with an Ordered Ter-Ter reaction mechanism [Marmor01].

MurC exists in both monomeric and dimeric form, and both are active [Jin96]. Crystal structures of the enzyme have been solved [Emanuele96, Deva03, Deva06].

Temperature-sensitive murC mutants have been isolated [Lugtenberg72, Miyakawa72]. The murC3 allele [Miyakawa72] was determined to carry a G344D mutation in a conserved region of the enzyme [Eveland97]. Additional point mutations were generated by site-directed mutagenesis; analysis of their effect on activity suggested the location of the active site [Bouhss97] and a residue involved in enzyme stability [Nosal98].

MurC is a potential antimicrobial drug target; inhibitors of MurC activity have been identified [Reck01, Marmor01, El03, Ehmann04, Frlan08, Zawadzke08, Sink08, Sova09, Rausch11, Hameed14].

Reviews: [Vicente98, El03a, Typas12]

Citations: [MenginLecreulx98, Liger96, Ikeda90, Zhang10b, Ishiguro82]

Gene Citations: [MenginLecreulx98, Hara97]

Locations: cytosol

Map Position: [100,765 -> 102,240] (2.17 centisomes)
Length: 1476 bp / 491 aa

Molecular Weight of Polypeptide: 53.626 kD (from nucleotide sequence), 50.0 kD (experimental) [Liger95 ]

pI: 5.84

Unification Links: ASAP:ABE-0000324 , CGSC:476 , DIP:DIP-10278N , EchoBASE:EB0614 , EcoGene:EG10619 , EcoliWiki:b0091 , Mint:MINT-1222662 , ModBase:P17952 , OU-Microarray:b0091 , PortEco:murC , PR:PRO_000023314 , Pride:P17952 , Protein Model Portal:P17952 , RefSeq:NP_414633 , RegulonDB:EG10619 , SMR:P17952 , String:511145.b0091 , UniProt:P17952

Relationship Links: InterPro:IN-FAMILY:IPR000713 , InterPro:IN-FAMILY:IPR004101 , InterPro:IN-FAMILY:IPR005758 , InterPro:IN-FAMILY:IPR013221 , InterPro:IN-FAMILY:IPR016040 , Panther:IN-FAMILY:PTHR23135:SF5 , PDB:Structure:1CC9 , PDB:Structure:2F00 , Pfam:IN-FAMILY:PF01225 , Pfam:IN-FAMILY:PF02875 , Pfam:IN-FAMILY:PF08245

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, Miyakawa72, Lugtenberg72]
GO:0007049 - cell cycle Inferred by computational analysis [UniProtGOA11]
GO:0008360 - regulation of cell shape Inferred by computational analysis [UniProtGOA11, GOA01]
GO:0009058 - biosynthetic process Inferred by computational analysis [GOA01]
GO:0051301 - cell division Inferred by computational analysis [UniProtGOA11, GOA01]
GO:0071555 - cell wall organization Inferred by computational analysis [UniProtGOA11]
Molecular Function: GO:0000287 - magnesium ion binding Inferred from experiment [Deva06]
GO:0005515 - protein binding Inferred from experiment [Rajagopala14, Deva06]
GO:0005524 - ATP binding Inferred from experiment Inferred by computational analysis [UniProtGOA11, GOA06, GOA01, Deva06]
GO:0008763 - UDP-N-acetylmuramate-L-alanine ligase activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01a, GOA01, Falk96, Liger95]
GO:0042803 - protein homodimerization activity Inferred from experiment [Jin96]
GO:0000166 - nucleotide binding Inferred by computational analysis [UniProtGOA11]
GO:0016874 - ligase activity Inferred by computational analysis [UniProtGOA11, GOA01]
Cellular Component: GO:0005737 - cytoplasm Inferred by computational analysis [UniProtGOA11a, UniProtGOA11, GOA06, GOA01]

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

Essentiality data for murC knockouts: ?

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

Credits:
Created 16-Jul-2013 by Keseler I , SRI International
Last-Curated ? 16-Jul-2013 by Keseler I , SRI International


Enzymatic reaction of: UDP-N-acetylmuramate-alanine ligase

Synonyms: UDP-N-acetylmuramoyl-L-alanine synthetase, UDP-N-acetylmuramate:L-alanine ligase (ADP-forming), L-alanine-adding enzyme

EC Number: 6.3.2.8

L-alanine + UDP-N-α-D-acetylmuramate + ATP <=> UDP-N-acetylmuramoyl-L-alanine + ADP + phosphate + H+

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 physiologically favored in the direction shown.

Alternative Substrates for L-alanine [Comment 2 ]: β-alanine [Emanuele96 ] , glycine [Liger95 ] , L-serine [Liger95 ]

Alternative Substrates for UDP-N-α-D-acetylmuramate [Comment 2 ]: beta-anomer of UDP-N-acetylmuramate [Liger95 ] , dihydro-UDP-N-acetylmuramate [Liger95 ] , UDP-N-acetyl-α-D-glucosamine-enolpyruvate [Liger95 ]

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

Summary:
It is possible to assay the enzymatic activity in the reverse direction; however, 10-fold more enzyme is required [Liger95].

Cofactors or Prosthetic Groups: Mn2+ [Emanuele96], Mg2+

Inhibitors (Unknown Mechanism): β-cyano-L-alanine [Liger95] , β-fluoro-L-alanine [Liger95] , 3-chloro-L-alanine [Liger95]

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
UDP-N-acetyl-α-D-glucosamine-enolpyruvate
430.0
[Liger95]
L-serine
850.0
[Liger95]
UDP-N-α-D-acetylmuramate
10.0
[Nosal98, BRENDA14]
UDP-N-α-D-acetylmuramate
34.3
[Deng04, BRENDA14]
UDP-N-α-D-acetylmuramate
39.0
[Falk96, BRENDA14]
UDP-N-α-D-acetylmuramate
44.0
[Emanuele96, BRENDA14]
UDP-N-α-D-acetylmuramate
58.0
[Gubler96, BRENDA14]
UDP-N-α-D-acetylmuramate
300.0
[MenginLecreulx82, BRENDA14]
UDP-N-α-D-acetylmuramate
100.0
[Liger96, BRENDA14]
UDP-N-α-D-acetylmuramate
100.0
[Liger95, BRENDA14]
dihydro-UDP-N-acetylmuramate
1000.0
[Liger95]
ATP
63.8
[Deng04, BRENDA14]
ATP
92.0
[Gubler96, BRENDA14]
ATP
130.0, 140.0
[Emanuele96, BRENDA14]
ATP
450.0
[Liger96, BRENDA14]
ATP
450.0
[Nosal98, BRENDA14]
ATP
450.0
[Liger95, BRENDA14]
β-alanine
46.0
2.5
[Emanuele96]
L-alanine
22.9
[Deng04, BRENDA14]
L-alanine
40.0
[Nosal98, BRENDA14]
L-alanine
20.0
[Liger95]
L-alanine
16.33
[Emanuele96]
glycine
2500.0
[Liger95]
beta-anomer of UDP-N-acetylmuramate
2000.0
[Liger95]

T(opt): 45 °C [BRENDA14, Liger95]

pH(opt) (forward direction): 8 [Emanuele96]

pH(opt): 8.6 [BRENDA14, Liger95], 9 [BRENDA14, Gubler96]


Sequence Features

Feature Class Location Citations Comment
Nucleotide-Phosphate-Binding-Region 126 -> 132
[UniProt10a]
UniProt: ATP; Non-Experimental Qualifier: potential;
Mutagenesis-Variant 344
[UniProt10]
Alternate sequence: G → D; UniProt: In murC3;


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
10/20/97 Gene b0091 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10619; 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

Bouhss97: Bouhss A, Mengin-Lecreulx D, Blanot D, van Heijenoort J, Parquet C (1997). "Invariant amino acids in the Mur peptide synthetases of bacterial peptidoglycan synthesis and their modification by site-directed mutagenesis in the UDP-MurNAc:L-alanine ligase from Escherichia coli." Biochemistry 36(39);11556-63. PMID: 9305945

Bouhss99: Bouhss A, Dementin S, van Heijenoort J, Parquet C, Blanot D (1999). "Formation of adenosine 5'-tetraphosphate from the acyl phosphate intermediate: a difference between the MurC and MurD synthetases of Escherichia coli." FEBS Lett 453(1-2);15-9. PMID: 10403366

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

Deng04: Deng G, Gu RF, Marmor S, Fisher SL, Jahic H, Sanyal G (2004). "Development of an LC-MS based enzyme activity assay for MurC: application to evaluation of inhibitors and kinetic analysis." J Pharm Biomed Anal 35(4);817-28. PMID: 15193726

Deva03: Deva T, Pryor KD, Leiting B, Baker EN, Smith CA (2003). "Purification, crystallization and preliminary X-ray analysis of Escherichia coli UDP-N-acetylmuramoyl:L-alanine ligase (MurC)." Acta Crystallogr D Biol Crystallogr 59(Pt 8);1510-3. PMID: 12876369

Deva06: Deva T, Baker EN, Squire CJ, Smith CA (2006). "Structure of Escherichia coli UDP-N-acetylmuramoyl:L-alanine ligase (MurC)." Acta Crystallogr D Biol Crystallogr 62(Pt 12);1466-74. PMID: 17139082

Ehmann04: Ehmann DE, Demeritt JE, Hull KG, Fisher SL (2004). "Biochemical characterization of an inhibitor of Escherichia coli UDP-N-acetylmuramyl-l-alanine ligase." Biochim Biophys Acta 1698(2);167-74. PMID: 15134649

El03: El Zoeiby A, Beaumont M, Dubuc E, Sanschagrin F, Voyer N, Levesque RC (2003). "Combinatorial enzymatic assay for the screening of a new class of bacterial cell wall inhibitors." Bioorg Med Chem 11(7);1583-92. PMID: 12628682

El03a: 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

Emanuele96: Emanuele JJ, Jin H, Jacobson BL, Chang CY, Einspahr HM, Villafranca JJ (1996). "Kinetic and crystallographic studies of Escherichia coli UDP-N-acetylmuramate:L-alanine ligase." Protein Sci 5(12);2566-74. PMID: 8976565

Eveland97: Eveland SS, Pompliano DL, Anderson MS (1997). "Conditionally lethal Escherichia coli murein mutants contain point defects that map to regions conserved among murein and folyl poly-gamma-glutamate ligases: identification of a ligase superfamily." Biochemistry 36(20);6223-9. PMID: 9166795

Falk96: Falk PJ, Ervin KM, Volk KS, Ho HT (1996). "Biochemical evidence for the formation of a covalent acyl-phosphate linkage between UDP-N-acetylmuramate and ATP in the Escherichia coli UDP-N-acetylmuramate:L-alanine ligase-catalyzed reaction." Biochemistry 35(5);1417-22. PMID: 8634271

Frlan08: Frlan R, Kovac A, Blanot D, Gobec S, Pecar S, Obreza A (2008). "Design and synthesis of novel N-benzylidenesulfonohydrazide inhibitors of MurC and MurD as potential antibacterial agents." Molecules 13(1);11-30. PMID: 18259126

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."

Gubler96: Gubler M, Appoldt Y, Keck W (1996). "Overexpression, purification, and characterization of UDP-N-acetylmuramyl:L-alanine ligase from Escherichia coli." J Bacteriol 178(3);906-10. PMID: 8550531

Hameed14: Hameed P S, Manjrekar P, Chinnapattu M, Humnabadkar V, Shanbhag G, Kedari C, Mudugal NV, Ambady A, de Jonge BL, Sadler C, Paul B, Sriram S, Kaur P, Guptha S, Raichurkar A, Fleming P, Eyermann CJ, McKinney DC, Sambandamurthy VK, Panda M, Ravishankar S (2014). "Pyrazolopyrimidines Establish MurC as a Vulnerable Target in Pseudomonas aeruginosa and Escherichia coli." ACS Chem Biol. PMID: 25035921

Hara97: Hara H, Yasuda S, Horiuchi K, Park JT (1997). "A promoter for the first nine genes of the Escherichia coli mra cluster of cell division and cell envelope biosynthesis genes, including ftsI and ftsW." J Bacteriol 179(18);5802-11. PMID: 9294438

Ikeda90: Ikeda M, Wachi M, Jung HK, Ishino F, Matsuhashi M (1990). "Nucleotide sequence involving murG and murC in the mra gene cluster region of Escherichia coli." Nucleic Acids Res 18(13);4014. PMID: 2197603

Ishiguro82: Ishiguro EE (1982). "Inhibition of uridine 5'-diphosphate-N-acetylmuramyl-L-alanine synthetase by beta-chloro-L-alanine in Escherichia coli." Can J Microbiol 28(6);654-9. PMID: 6811120

Ito73: Ito E, Strominger JL (1973). "Enzymatic synthesis of the peptide in bacterial uridine nucleotides. VII. Comparative biochemistry." J Biol Chem 248(9);3131-6. PMID: 4633672

Jin96: Jin H, Emanuele JJ, Fairman R, Robertson JG, Hail ME, Ho HT, Falk PJ, Villafranca JJ (1996). "Structural studies of Escherichia coli UDP-N-acetylmuramate:L-alanine ligase." Biochemistry 35(5);1423-31. PMID: 8634272

Liger91: Liger D, Blanot D, van Heijenoort J (1991). "Effect of various alanine analogues on the L-alanine-adding enzyme from Escherichia coli." FEMS Microbiol Lett 64(1);111-5. PMID: 1855644

Liger95: Liger D, Masson A, Blanot D, van Heijenoort J, Parquet C (1995). "Over-production, purification and properties of the uridine-diphosphate-N-acetylmuramate:L-alanine ligase from Escherichia coli." Eur J Biochem 1995;230(1);80-7. PMID: 7601127

Liger96: Liger D, Masson A, Blanot D, van Heijenoort J, Parquet C (1996). "Study of the overproduced uridine-diphosphate-N-acetylmuramate:L-alanine ligase from Escherichia coli." Microb Drug Resist 2(1);25-7. PMID: 9158719

Lugtenberg72: Lugtenberg EJ, v Schijndel-van Dam A (1972). "Temperature-sensitive mutants of Escherichia coli K-12 with low activities of the L-alanine adding enzyme and the D-alanyl-D-alanine adding enzyme." J Bacteriol 110(1);35-40. PMID: 4552998

Lugtenberg72a: Lugtenberg EJ (1972). "Studies on Escherichia coli enzymes involved in the synthesis of uridine diphosphate-N-acetyl-muramyl-pentapeptide." J Bacteriol 110(1);26-34. PMID: 4552992

Marmor01: Marmor S, Petersen CP, Reck F, Yang W, Gao N, Fisher SL (2001). "Biochemical characterization of a phosphinate inhibitor of Escherichia coli MurC." Biochemistry 40(40);12207-14. PMID: 11580296

MenginLecreulx82: Mengin-Lecreulx D, Flouret B, van Heijenoort J (1982). "Cytoplasmic steps of peptidoglycan synthesis in Escherichia coli." J Bacteriol 151(3);1109-17. PMID: 6125497

MenginLecreulx98: Mengin-Lecreulx D, Ayala J, Bouhss A, van Heijenoort J, Parquet C, Hara H (1998). "Contribution of the Pmra promoter to expression of genes in the Escherichia coli mra cluster of cell envelope biosynthesis and cell division genes." J Bacteriol 180(17);4406-12. PMID: 9721276

Miyakawa72: Miyakawa T, Matsuzawa H, Matsuhashi M, Sugino Y (1972). "Cell wall peptidoglycan mutants of Escherichia coli K-12: existence of two clusters of genes, mra and mrb, for cell wall peptidoglycan biosynthesis." J Bacteriol 112(2);950-8. PMID: 4563986

Nosal98: Nosal F, Masson A, Legrand R, Blanot D, Schoot B, van Heijenoort J, Parquet C (1998). "Site-directed mutagenesis and chemical modification of the two cysteine residues of the UDP-N-acetylmuramoyl:L-alanine ligase of Escherichia coli." FEBS Lett 426(3);309-13. PMID: 9600257

Pryor97: Pryor KD, Leiting B (1997). "High-level expression of soluble protein in Escherichia coli using a His6-tag and maltose-binding-protein double-affinity fusion system." Protein Expr Purif 10(3);309-19. PMID: 9268677

Rajagopala14: Rajagopala SV, Sikorski P, Kumar A, Mosca R, Vlasblom J, Arnold R, Franca-Koh J, Pakala SB, Phanse S, Ceol A, Hauser R, Siszler G, Wuchty S, Emili A, Babu M, Aloy P, Pieper R, Uetz P (2014). "The binary protein-protein interaction landscape of Escherichia coli." Nat Biotechnol 32(3);285-90. PMID: 24561554

Rausch11: Rausch S, Hanchen A, Denisiuk A, Lohken M, Schneider T, Sussmuth RD (2011). "Feglymycin is an inhibitor of the enzymes MurA and MurC of the peptidoglycan biosynthesis pathway." Chembiochem 12(8);1171-3. PMID: 21538763

Reck01: Reck F, Marmor S, Fisher S, Wuonola MA (2001). "Inhibitors of the bacterial cell wall biosynthesis enzyme MurC." Bioorg Med Chem Lett 11(11);1451-4. PMID: 11378375

Sink08: Sink R, Kovac A, Tomasic T, Rupnik V, Boniface A, Bostock J, Chopra I, Blanot D, Masic LP, Gobec S, Zega A (2008). "Synthesis and biological evaluation of N-acylhydrazones as inhibitors of MurC and MurD ligases." ChemMedChem 3(9);1362-70. PMID: 18651694

Sova09: Sova M, Kovac A, Turk S, Hrast M, Blanot D, Gobec S (2009). "Phosphorylated hydroxyethylamines as novel inhibitors of the bacterial cell wall biosynthesis enzymes MurC to MurF." Bioorg Chem 37(6);217-22. PMID: 19804894

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.

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 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."

Vicente98: Vicente M, Gomez MJ, Ayala JA (1998). "Regulation of transcription of cell division genes in the Escherichia coli dcw cluster." Cell Mol Life Sci 54(4);317-24. PMID: 9614967

Zawadzke08: Zawadzke LE, Norcia M, Desbonnet CR, Wang H, Freeman-Cook K, Dougherty TJ (2008). "Identification of an inhibitor of the MurC enzyme, which catalyzes an essential step in the peptidoglycan precursor synthesis pathway." Assay Drug Dev Technol 6(1);95-103. PMID: 18315498

Zhang10b: Zhang X, El-Hajj ZW, Newman E (2010). "Deficiency in L-serine deaminase interferes with one-carbon metabolism and cell wall synthesis in Escherichia coli K-12." J Bacteriol 192(20);5515-25. PMID: 20729359

Other References Related to Gene Regulation

Eraso14: Eraso JM, Markillie LM, Mitchell HD, Taylor RC, Orr G, Margolin W (2014). "The Highly Conserved MraZ Protein Is a Transcriptional Regulator in Escherichia coli." J Bacteriol 196(11);2053-66. PMID: 24659771

Gohler11: Gohler AK, Kokpinar O, Schmidt-Heck W, Geffers R, Guthke R, Rinas U, Schuster S, Jahreis K, Kaleta C (2011). "More than just a metabolic regulator - elucidation and validation of new targets of PdhR in Escherichia coli." BMC Syst Biol 5(1);197. PMID: 22168595

Ishino89: Ishino F, Jung HK, Ikeda M, Doi M, Wachi M, Matsuhashi M (1989). "New mutations fts-36, lts-33, and ftsW clustered in the mra region of the Escherichia coli chromosome induce thermosensitive cell growth and division." J Bacteriol 171(10);5523-30. PMID: 2676977


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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 Sun Nov 23, 2014, biocyc13.