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Literature summary for 6.3.2.7 extracted from

  • Ruane, K.M.; Lloyd, A.J.; Fueloep, V.; Dowson, C.G.; Barreteau, H.; Boniface, A.; Dementin, S.; Blanot, D.; Mengin-Lecreulx, D.; Gobec, S.; Dessen, A.; Roper, D.I.
    Specificity determinants for lysine incorporation in Staphylococcus aureus peptidoglycan as revealed by the structure of a MurE enzyme ternary complex (2013), J. Biol. Chem., 288, 33439-33448.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene murE, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3):pLysS from plasmid pET2160:murESa, coexpression with chaperone from pREP4groESL Staphylococcus aureus

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant His-tagged enzyme in complex with UDP-MurNAc-Ala-Glu-Lys or with UDP-MurNAc-Ala-Glu-Lys and ADP, from 0.1 M Na-HEPES/MOPS, pH 7.5, 0.09 M nitrate phosphate sulfate mix, 30% w/v PEG550MME/PEG20K, method evaluation, X-ray diffraction structure determination and analysis at 1.8-1.9 A resolution, molecular replacement Staphylococcus aureus

Protein Variants

Protein Variants Comment Organism
A409R site-directed mutagenesis, the lytic phenotype observed upon overexpression of the wild-type murESa gene is replicated with the mutant Staphylococcus aureus
D406A site-directed mutagenesis, the mutant shows very low or no activity in vivo Staphylococcus aureus
E460A site-directed mutagenesis, the mutant shows very low or no activity in vivo Staphylococcus aureus
N407A site-directed mutagenesis, inactive mutant Staphylococcus aureus
N407R site-directed mutagenesis, the growth curve of N407R mutant is similar to that of wild-type murESa Staphylococcus aureus
P408A site-directed mutagenesis, the lytic phenotype observed upon overexpression of the wild-type murESa gene is replicated with the mutant Staphylococcus aureus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.55
-
L-lysine pH 8.6, 37°C, recombinant wild-type enzyme Staphylococcus aureus
2.8
-
L-lysine pH 8.6, 37°C, recombinant mutant D406A Staphylococcus aureus
3.1
-
L-lysine pH 8.6, 37°C, recombinant mutant P408A Staphylococcus aureus
20
-
L-lysine above, pH 8.6, 37°C, recombinant mutant A409R Staphylococcus aureus
20
-
L-lysine above, pH.6, 37°C, recombinant mutant E460A Staphylococcus aureus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Staphylococcus aureus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + UDP-N-acetylmuramoyl-L-alanyl-D-glutamate + L-lysine Staphylococcus aureus the enzyme of the Gram-positive bacterium discriminates between L-lysine and D,L-diaminopimelic acid, the predominant amino acid that replaces L-lysine in Gram-negative peptidoglycan. Despite the absolute specificity for L-lysine, Staphylococcus aureus MurE binds this substrate relatively poorly. In vivo analysis and metabolomic data reveal that this is compensated for by high cytoplasmic L-lysine concentrations ADP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysine
-
?
additional information Staphylococcus aureus the third amino acid added to the peptide stem of peptidoglycan is usually either meso-diaminopimelic acid (in most Gram-negative bacteria), EC 6.3.2.13, or L-lysine (in most Gram-positive bacteria), EC 6.3.2.7 ?
-
?

Organism

Organism UniProt Comment Textmining
Staphylococcus aureus Q2FZP6
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3):pLysS by nickel affinity chromatography, gel filtration, and ion exchange chromatography Staphylococcus aureus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + UDP-N-acetylmuramoyl-L-alanyl-D-glutamate + L-lysine the enzyme of the Gram-positive bacterium discriminates between L-lysine and D,L-diaminopimelic acid, the predominant amino acid that replaces L-lysine in Gram-negative peptidoglycan. Despite the absolute specificity for L-lysine, Staphylococcus aureus MurE binds this substrate relatively poorly. In vivo analysis and metabolomic data reveal that this is compensated for by high cytoplasmic L-lysine concentrations Staphylococcus aureus ADP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysine
-
?
ATP + UDP-N-acetylmuramoyl-L-alanyl-D-glutamate + L-lysine despite the absolute specificity for L-lysine, Staphylococcus aureus MurE binds this substrate relatively poorly. In vivo analysis and metabolomic data reveal that this is compensated for by high cytoplasmic L-lysine concentrations, substrate discrimination mechanism, overview. Glu460 is crucial for interaction with the epsilon-amino group of the L-lysine substrate Staphylococcus aureus ADP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysine
-
?
additional information the third amino acid added to the peptide stem of peptidoglycan is usually either meso-diaminopimelic acid (in most Gram-negative bacteria), EC 6.3.2.13, or L-lysine (in most Gram-positive bacteria), EC 6.3.2.7 Staphylococcus aureus ?
-
?

Synonyms

Synonyms Comment Organism
MurE
-
Staphylococcus aureus
UDP-N-acetylmuramoyl-L-alanyl-D-glutamate:meso-2,6-diaminopimelate ligase
-
Staphylococcus aureus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Staphylococcus aureus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.001
-
L-lysine pH 8.6, 37°C, recombinant mutant D406A Staphylococcus aureus
0.283
-
L-lysine pH 8.6, 37°C, recombinant mutant P408A Staphylococcus aureus
4.833
-
L-lysine pH 8.6, 37°C, recombinant wild-type enzyme Staphylococcus aureus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.6
-
assay at Staphylococcus aureus

Cofactor

Cofactor Comment Organism Structure
ATP
-
Staphylococcus aureus

General Information

General Information Comment Organism
metabolism the enzyme is involved in peptidoglycan biosynthesis and cell wall assembly Staphylococcus aureus
additional information ATP-binding site structure and homology, overview. Asn407 makes a series of structurally important hydrogen bond interactions within the context of the architecture of the active site region Staphylococcus aureus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.001
-
L-lysine pH 8.6, 37°C, recombinant mutant D406A Staphylococcus aureus
0.003
-
L-lysine above, pH 8.6, 37°C, recombinant mutant A409R Staphylococcus aureus
0.029
-
L-lysine above, pH.6, 37°C, recombinant mutant E460A Staphylococcus aureus
0.091
-
L-lysine pH 8.6, 37°C, recombinant mutant P408A Staphylococcus aureus
8.787
-
L-lysine pH 8.6, 37°C, recombinant wild-type enzyme Staphylococcus aureus