Any feedback?
Please rate this page
(enzyme.php)
(0/150)

BRENDA support

BRENDA Home
show all | hide all No of entries

Information on EC 6.3.2.10 - UDP-N-acetylmuramoyl-tripeptide-D-alanyl-D-alanine ligase and Organism(s) Escherichia coli and UniProt Accession P11880

for references in articles please use BRENDA:EC6.3.2.10
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
IUBMB Comments
Involved with EC 6.3.2.4 (D-alanine---D-alanine ligase), EC 6.3.2.7 (UDP-N-acetylmuramoyl-L-alanyl-D-glutamate---L-lysine ligase) or EC 6.3.2.13 (UDP-N-acetylmuramoyl-L-alanyl-D-glutamate---2,6-diaminopimelate ligase), EC 6.3.2.8 (UDP-N-acetylmuramate---L-alanine ligase) and EC 6.3.2.9 (UDP-N-acetylmuramoyl-L-alanine---D-glutamate ligase) in the synthesis of a cell-wall peptide (click here) for diagram. This enzyme also catalyses the reaction when the C-terminal residue of the tripeptide is meso-2,6-diaminoheptanedioate (acylated at its L-centre), linking the D-Ala-D-Ala to the carboxy group of the L-centre. This activity was previously attributed to EC 6.3.2.15, which has since been deleted.
Specify your search results
Select one or more organisms in this record: ?
This record set is specific for:
Escherichia coli
UNIPROT: P11880
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)
Word Map
hide
The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Archaea
Synonyms
murf1, d-ala-d-ala-adding enzyme, abmurf, atp-dependent udp-n-acetylmuramoyl-tripeptide-d-alanyl-d-alanine ligase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D-Ala-D-Ala-adding enzyme
-
Synthetase, uridine diphosphoacetylmuramoylpentapeptide
-
UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine synthetase
-
UDP-N-acetylmuramoylalanyl-D-glutamyl-lysine-D-alanyl-D-alanine ligase
-
UDPacetylmuramoylpentapeptide synthetase
-
Uridine diphosphoacetylmuramoylpentapeptide synthetase
-
D-Ala-D-Ala-adding enzyme
Synthetase, uridine diphosphoacetylmuramoylpentapeptide
UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine synthetase
UDP-N-acetylmuramoylalanyl-D-glutamyl-lysine-D-alanyl-D-alanine ligase
UDPacetylmuramoylpentapeptide synthetase
Uridine diphosphoacetylmuramoylpentapeptide synthetase
additional information
member of ligase superfamily
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + UDP-N-acetylmuramoyl-L-alanyl-gamma-D-glutamyl-L-lysine + D-alanyl-D-alanine = ADP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-gamma-D-glutamyl-L-lysyl-D-alanyl-D-alanine
show the reaction diagram
ordered kinetic mechanism, E158 and H188 are critical for enzyme activity
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
carboxylic acid amide formation
-
carboxamide formation
-
carboxylic acid amide formation
carboxamide formation
PATHWAY SOURCE
PATHWAYS
-
-, -, -
SYSTEMATIC NAME
IUBMB Comments
UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysine:D-alanyl-D-alanine ligase (ADP-forming)
Involved with EC 6.3.2.4 (D-alanine---D-alanine ligase), EC 6.3.2.7 (UDP-N-acetylmuramoyl-L-alanyl-D-glutamate---L-lysine ligase) or EC 6.3.2.13 (UDP-N-acetylmuramoyl-L-alanyl-D-glutamate---2,6-diaminopimelate ligase), EC 6.3.2.8 (UDP-N-acetylmuramate---L-alanine ligase) and EC 6.3.2.9 (UDP-N-acetylmuramoyl-L-alanine---D-glutamate ligase) in the synthesis of a cell-wall peptide (click here) for diagram. This enzyme also catalyses the reaction when the C-terminal residue of the tripeptide is meso-2,6-diaminoheptanedioate (acylated at its L-centre), linking the D-Ala-D-Ala to the carboxy group of the L-centre. This activity was previously attributed to EC 6.3.2.15, which has since been deleted.
CAS REGISTRY NUMBER
COMMENTARY hide
9023-60-3
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + UDP-N-acetylmuramoyl-L-Ala-D-Glu-L-Lys + D-Ala-D-Ala
ADP + phosphate + UDP-N-acetylmuramoyl-L-Ala-D-Glu-L-Lys-D-Ala-D-Ala
show the reaction diagram
ATP + UDP-N-acetylmuramoyl-L-Ala-D-Glu-L-Lys + D-Ala-D-Ala
ADP + phosphate + UDP-N-acetylmuramoyl-L-Ala-D-Glu-L-Lys-D-Ala-D-Ala
show the reaction diagram
ATP + UDP-N-acetylmuramoyl-L-Ala-D-Glu-L-Lys + fluoro-D-Ala-fluoro-D-Ala
ADP + phosphate + UDP-N-acetylmuramoyl-L-Ala-D-Glu-L-Lys-fluoro-D-Ala-fluoro-D-Ala
show the reaction diagram
-
-
-
?
ATP + UDP-N-acetylmuramoyl-L-alanyl-gamma-D-glutamyl-diaminopimelic acid + D-alanyl-D-alanine
ADP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-gamma-D-glutamyl-diaminopimeloyl-D-alanyl-D-alanine
show the reaction diagram
-
-
-
-
?
ATP + UDP-N-acetylmuramoyl-L-alanyl-gamma-D-glutamyl-L-lysine + D-alanyl-D-alanine
ADP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-gamma-D-glutamyl-L-lysyl-D-alanyl-D-alanine
show the reaction diagram
additional information
?
-
the specific activity of the mutant enzyme is highly reduced compared to the wild-type murF
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + UDP-N-acetylmuramoyl-L-Ala-D-Glu-L-Lys + D-Ala-D-Ala
ADP + phosphate + UDP-N-acetylmuramoyl-L-Ala-D-Glu-L-Lys-D-Ala-D-Ala
show the reaction diagram
ATP + UDP-N-acetylmuramoyl-L-Ala-D-Glu-L-Lys + D-Ala-D-Ala
ADP + phosphate + UDP-N-acetylmuramoyl-L-Ala-D-Glu-L-Lys-D-Ala-D-Ala
show the reaction diagram
-
-
-
?
ATP + UDP-N-acetylmuramoyl-L-alanyl-gamma-D-glutamyl-L-lysine + D-alanyl-D-alanine
ADP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-gamma-D-glutamyl-L-lysyl-D-alanyl-D-alanine
show the reaction diagram
-
MurF catalyzes the last cytoplasmic step of bacterial cell wall synthesis
-
-
?
additional information
?
-
the specific activity of the mutant enzyme is highly reduced compared to the wild-type murF
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
-
required
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2R,4R)-4-(3,7-dimethoxynaphthalen-2-yl)-1-(dimethylamino)-3,4-diphenylbutan-2-ol
-
modest activity against lipopolysaccharide-defective Escherichia coli and wild-type Escherichia coli rendered permeable with polymyxin B nonapeptide. Treatment of lipopolysaccharide-defective Escherichia coli cells with >2fold minimal inhibitory concentrations of DQ1 results in a 75-fold-greater accumulation of the MurF substrate compared to the control, a 70% decline in the amount of the MurF product, and eventual cell lysis, consistent with the inhibition of MurF within bacteria
(2S,4R)-4-(3,7-dimethoxynaphthalen-2-yl)-1-(dimethylamino)-3,4-diphenylbutan-2-ol
-
additionally active against Gram-positive bacteria, including methicillin-susceptible and -resistant Staphylococcus aureus isolates and vancomycin-susceptible and -resistant Enterococcus faecalis and Enterococcus faecium isolates
1-[[(4-trifluoromethylphenyl)sulfonyl]amino]-3-(morpholin-4-yl)propan-2-yl dihydrogen phosphate
-
-
4-[(pentachlorobenzyl)sulfanyl]-6-sulfanyl-1,3,5-triazin-2-ol
-
i.e. NSC 209931, inhibitor identified by structure-based virtual screening
acetate
-
inhibition of wild-type enzyme
Butylamine
Butyrate
-
slight inhibition of wild-type enzyme
ethylamine
-
inhibition of wild-type enzyme
formate
-
inhibition of wild-type enzyme
methylamine
-
inhibition of wild-type enzyme
N-(3-aminopropyl)-4-(trifluoromethyl)-2-([4-[3-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl]methyl)pyrimidine-5-carboxamide
-
-
N-(4-carbamimidoylphenyl)-4-(trifluoromethyl)-2-([4-[3-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl]methyl)pyrimidine-5-carboxamide
-
-
N-[3-(cyclopentylamino)propyl]-2-[[4-(4-methylphenyl)-1,3-thiazol-2-yl]amino]-4-(trifluoromethyl)pyrimidine-5-carboxamide
-
-
propionate
-
inhibition of wild-type enzyme
Propylamine
-
inhibition of wild-type enzyme
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
acetate
-
activation of mutant K202A
Butyrate
-
activation of mutant K202A
ethylamine
-
slight activation of mutant K202A
formate
-
slight activation of mutant K202A
propionate
-
strong activation of mutant K202A
Propylamine
-
slight activation of mutant K202A
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
-
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.024
(2R,4R)-4-(3,7-dimethoxynaphthalen-2-yl)-1-(dimethylamino)-3,4-diphenylbutan-2-ol
Escherichia coli
-
-
0.15
1-[[(4-trifluoromethylphenyl)sulfonyl]amino]-3-(morpholin-4-yl)propan-2-yl dihydrogen phosphate
Escherichia coli
-
pH 8.0
0.063
4-[(pentachlorobenzyl)sulfanyl]-6-sulfanyl-1,3,5-triazin-2-ol
Escherichia coli
-
pH 8.0, 37°C
0.0075
N-(3-aminopropyl)-4-(trifluoromethyl)-2-([4-[3-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl]methyl)pyrimidine-5-carboxamide
Escherichia coli
-
-
0.0025
N-(4-carbamimidoylphenyl)-4-(trifluoromethyl)-2-([4-[3-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl]methyl)pyrimidine-5-carboxamide
Escherichia coli
-
-
0.0025
N-[3-(cyclopentylamino)propyl]-2-[[4-(4-methylphenyl)-1,3-thiazol-2-yl]amino]-4-(trifluoromethyl)pyrimidine-5-carboxamide
Escherichia coli
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.006
-
purified recombinant mutant K202A, in presence of butylamine
0.016
-
purified recombinant mutant K202A
0.032
-
purified recombinant mutant K202A, in presence of propylamine
0.046
-
purified recombinant mutant K202A, in presence of ethylamine
0.052
-
purified recombinant mutant K202A, in presence of formate
0.062
0.437
-
purified recombinant mutant K202A, in presence of butyrate
0.511
-
purified recombinant mutant K202A, in presence of acetate
0.55
-
purified recombinant wild-type enzyme, in presence of methylamine
0.71
-
purified recombinant wild-type enzyme, in presence of propylamine
0.72
-
purified recombinant wild-type enzyme, in presence of butylamine
0.76
-
purified recombinant wild-type enzyme, in presence of propionate
0.769
-
purified recombinant mutant K202A, in presence of propionate
0.8
-
purified recombinant wild-type enzyme, in presence of ethylamine
0.88
-
purified recombinant wild-type enzyme, in presence of acetate
0.93
-
purified recombinant wild-type enzyme, in presence of formate
1.24
-
purified recombinant wild-type enzyme, in presence of butyrate
1.27
-
purified recombinant wild-type enzyme
11.2
purified recombinant wild-type enzyme, 30°C
13.9
purified recombinant wild-type enzyme, 42°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.6
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
assay at
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 4792, nucleotide sequence
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging-drop vapour diffusion method, purified recombinant enzyme 12.2 mg/ml, in 10 mM Tris-HCl, pH 7.4, 10 mM DTT, plus equal volume of reservoir solution: 0.1 M Bis-Tris propane buffer, pH 9.4, 18% PEG 8000, 0.12 M MgSO4, 10% glycerol, about 20 days, X-ray diffraction structure determination at 2.8 A resolution, and analysis
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A288T
naturally occuring murF2 allele, expressed as a fusion protein with glutathione S-transferase, is 181fold less catalytically active than the wild-type enzyme at 30°c and even more reduced at 42°C
E158A
site-directed mutagenesis, 4520fold reduced activity compared to the wild-type
E158D
site-directed mutagenesis, 246fold reduced activity compared to the wild-type
E158G
site-directed mutagenesis, over 451fold reduced activity compared to the wild-type
H188A
site-directed mutagenesis, 203fold reduced activity compared to the wild-type
H188D
site-directed mutagenesis, 2990fold reduced activity compared to the wild-type
H188G
site-directed mutagenesis, 214fold reduced activity compared to the wild-type
H188N
site-directed mutagenesis, 1860fold reduced activity compared to the wild-type
K202A
-
site-directed mutagenesis, exchange of highly conserved lysine residue leads to highly reduced activity, activity can be rescued best by addition of propionate or other short-chain carboxylic acids, but only in a small extent by amines
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant from strain BL21(DE3)
recombinant wild-type and mutant enzyme from strain JM83
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in strain BL21(DE3)
expression of naturally occuring mutant allele murF2 with A288T mutation as glutathione S-transferase fusion protein
expression of point mutants, expression of wild-type and naturally occuring mutant allele murF2 with A288T mutation as glutathione S-transferase fusion proteins
overexpression of wild-type and MurF mutant K202A as His-tagged enzymes in strain JM83
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pharmacology
attractive target for development of antibacterial agents
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Parquet, C.; Flouret, B.; Mengin-Lecreulx, D.; van Heijenoort, J.
Nucleotide sequence of the murF gene encoding the UDP-MurNAc-pentapeptide synthetase of Escherichia coli
Nucleic Acids Res.
17
5379
1989
Escherichia coli
Manually annotated by BRENDA team
Walsh, C.T.
Enzymes in the D-alanine branch of bacterial cell wall peptidoglycan assembly
J. Biol. Chem.
264
2393-2396
1989
Escherichia coli
Manually annotated by BRENDA team
Yan, Y.; Munshi, S.; Li, Y.; Pryor, K.A.; Marsilio, F.; Leiting, B.
Crystallization and preliminary X-ray analysis of the Escherichia coli UDP-MurNAc-tripeptide D-alanyl-D-alanine-adding enzyme (MurF)
Acta Crystallogr. Sect. D
55
2033-2034
1999
Escherichia coli (P11880), Escherichia coli K37 (P11880)
Manually annotated by BRENDA team
Eveland, S.S.; Pompliano, D.L.; Anderson, M.S.
Conditionally lethal Escherichia coli murein mutants contain point defects that map to regions conserved among murein and folyl poly-g-glutamate ligases: identification of a ligase superfamily
Biochemistry
36
6223-6229
1997
Escherichia coli (P11880)
Manually annotated by BRENDA team
Dementin, S.; Bouhss, A.; Auger, G.; Parquet, C.; Mengin-Lecreulx, D.; Dideberg, O.; van Heijenoort, J.; Blanot, D.
Evidence of a functional requirement for a carbamoylated lysine residue in MurD, MurE and MurF synthetases as established by chemical rescue experiments
Eur. J. Biochem.
268
5800-5807
2001
Escherichia coli
Manually annotated by BRENDA team
Baum, E.Z.; Crespo-Carbone, S.M.; Abbanat, D.; Foleno, B.; Maden, A.; Goldschmidt, R.; Bush, K.
Utility of muropeptide ligase for identification of inhibitors of the cell wall biosynthesis enzyme MurF
Antimicrob. Agents Chemother.
50
230-236
2006
Escherichia coli
Manually annotated by BRENDA team
Baum, E.Z.; Crespo-Carbone, S.M.; Foleno, B.D.; Simon, L.D.; Guillemont, J.; Macielag, M.; Bush, K.
MurF inhibitors with antibacterial activity: effect on muropeptide levels
Antimicrob. Agents Chemother.
53
3240-3247
2009
Escherichia coli
Manually annotated by BRENDA team
Sova, M.; Kovac, A.; Turk, S.; Hrast, M.; Blanot, D.; Gobec, S.
Phosphorylated hydroxyethylamines as novel inhibitors of the bacterial cell wall biosynthesis enzymes MurC to MurF
Bioorg. Chem.
37
217-222
2009
Escherichia coli
Manually annotated by BRENDA team
Turk, S.; Kovac, A.; Boniface, A.; Bostock, J.M.; Chopra, I.; Blanot, D.; Gobec, S.
Discovery of new inhibitors of the bacterial peptidoglycan biosynthesis enzymes MurD and MurF by structure-based virtual screening
Bioorg. Med. Chem.
17
1884-1889
2009
Escherichia coli
Manually annotated by BRENDA team