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Literature summary for 6.3.2.13 extracted from

  • Basavannacharya, C.; Moody, P.R.; Munshi, T.; Cronin, N.; Keep, N.H.; Bhakta, S.
    Essential residues for the enzyme activity of ATP-dependent MurE ligase from Mycobacterium tuberculosis (2010), Protein Cell, 1, 1011-1022.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a His-tagged fusion protein Mycobacterium tuberculosis

Protein Variants

Protein Variants Comment Organism
D392A kcat lower than wild-type, Km (meso-2,6-diaminoheptanedioate) and Km (ATP) higher than wild-type, Km (UDP-MurNAc-L-Ala-D-Glu) lower than wild-type, mutant shows hydrolysis of ATP uncoupled from catalysis. The ATP hydrolysis rate is enhanced by at least partial occupation of the uridine nucleotide dipeptide binding site, mutant shows very high activity with all the amino acids, uridine sugar precursors and nucleotides tested compared to wild type and moreover, reaching up to 100% Mycobacterium tuberculosis
DELTA1-24 N-terminal deletion mutant, kcat comparable to wild-type, Km values comparable to wild-type, mutant shows similar results to the wild-type with all the nucleotides, UDP-MurNAc peptides and amino acids Mycobacterium tuberculosis
E220A kcat lower than wild-type, Km (ATP) and Km (UDP-MurNAc-L-Ala-D-Glu), Km (meso-2,6-diaminoheptanedioate) lower than wild-type, mutant shows hydrolysis of ATP uncoupled from catalysis. The ATP hydrolysis rate is enhanced by atleast partial occupation of the uridine nucleotide dipeptide binding site, mutant shows very high activity with all the amino acids, uridine sugar precursors and nucleotides tested compared to wild type and moreover, reaching up to 100% Mycobacterium tuberculosis
K157A kcat lower than wild-type, Km (meso-2,6-diaminoheptanedioate), Km (UDP-MurNAc-L-Ala-D-Glu) and Km (ATP) lower than wild-type, mutant shows hydrolysis of ATP uncoupled from catalysis. The ATP hydrolysis rate is enhanced by at least partial occupation of the uridine nucleotide dipeptide binding site, mutant shows very high activity with all the amino acids, uridine sugar precursors and nucleotides tested compared to wild type and moreover, reaching up to 100% Mycobacterium tuberculosis
N449D kcat lower than wild-type, Km (ATP) and Km (meso-2,6-diaminoheptanedioate) higher than wild-type, Km (UDP-MurNAc-L-Ala-D-Glu) lower than wild-type, mutant shows similar results to the wild-type with all the nucleotides, UDP-MurNAc peptides and amino acids, except D-Lys, which shows a slightly higher activity for mutant N449D Mycobacterium tuberculosis
R451A kcat lower than wild-type, Km (ATP) comparable to wild-type, Km (meso-2,6-diaminoheptanedioate) higher than wild-type, Km (UDP-MurNAc-L-Ala-D-Glu) lower than wild-type, significant activity in the presence of L-Lys, D-Lys, DL-ornithine and D-Glu for mutant R451A with similar activities as wild type for all the nucleotides and UDP-MurNAc peptides tested Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.005
-
UDP-MurNAc-L-Ala-D-Glu mutant K157A, pH 8.5, 37°C, value below Mycobacterium tuberculosis
0.005
-
UDP-MurNAc-L-Ala-D-Glu mutant R451A, pH 8.5, 37°C Mycobacterium tuberculosis
0.007
-
UDP-MurNAc-L-Ala-D-Glu mutant N449D, pH 8.5, 37°C Mycobacterium tuberculosis
0.008
-
ATP mutant K157A, pH 8.5, 37°C Mycobacterium tuberculosis
0.012
-
meso-2,6-diaminoheptanedioate mutant K157A, pH 8.5, 37°C Mycobacterium tuberculosis
0.015
-
ATP mutant R451A, pH 8.5, 37°C Mycobacterium tuberculosis
0.016
-
UDP-MurNAc-L-Ala-D-Glu mutant D392A, pH 8.5, 37°C Mycobacterium tuberculosis
0.017
-
ATP wild-type, pH 8.5, 37°C Mycobacterium tuberculosis
0.019
-
UDP-MurNAc-L-Ala-D-Glu DELTA1-24 (N-terminal deletion mutant), pH 8.5, 37°C Mycobacterium tuberculosis
0.021
-
ATP DELTA1-24 (N-terminal deletion mutant), pH 8.5, 37°C Mycobacterium tuberculosis
0.024
-
UDP-MurNAc-L-Ala-D-Glu wild-type, pH 8.5, 37°C Mycobacterium tuberculosis
0.052
-
meso-2,6-diaminoheptanedioate mutant E220A, pH 8.5, 37°C Mycobacterium tuberculosis
0.06
-
ATP mutant N449D, pH 8.5, 37°C Mycobacterium tuberculosis
0.065
-
meso-2,6-diaminoheptanedioate DELTA1-24 (N-terminal deletion mutant), pH 8.5, 37°C Mycobacterium tuberculosis
0.078
-
meso-2,6-diaminoheptanedioate wild-type, pH 8.5, 37°C Mycobacterium tuberculosis
0.1
-
ATP mutant D392A, pH 8.5, 37°C Mycobacterium tuberculosis
0.113
-
meso-2,6-diaminoheptanedioate mutant D392A, pH 8.5, 37°C Mycobacterium tuberculosis
0.123
-
ATP mutant E220A, pH 8.5, 37°C Mycobacterium tuberculosis
0.148
-
UDP-MurNAc-L-Ala-D-Glu mutant E220A, pH 8.5, 37°C Mycobacterium tuberculosis
0.324
-
meso-2,6-diaminoheptanedioate mutant N449D, pH 8.5, 37°C Mycobacterium tuberculosis
0.473
-
meso-2,6-diaminoheptanedioate mutant R451A, pH 8.5, 37°C Mycobacterium tuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Mycobacterium tuberculosis

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + MurNAc-L-Ala-D-Glu + meso-2,6-diaminoheptanedioate relative activity wild-type: 14% Mycobacterium tuberculosis ?
-
?
ATP + UDP-MurNAc-L-Ala-D-Glu + D-Ala-D-Ala relative activity wild-type: 5% Mycobacterium tuberculosis ?
-
?
ATP + UDP-MurNAc-L-Ala-D-Glu + D-Glu relative activity wild-type: 13% Mycobacterium tuberculosis ?
-
?
ATP + UDP-MurNAc-L-Ala-D-Glu + D-Lys relative activity wild-type: 9% Mycobacterium tuberculosis ?
-
?
ATP + UDP-MurNAc-L-Ala-D-Glu + D-ornithine relative activity wild-type: 8% Mycobacterium tuberculosis ?
-
?
ATP + UDP-MurNAc-L-Ala-D-Glu + DL-lanthionine relative activity wild-type: 82% Mycobacterium tuberculosis ?
-
?
ATP + UDP-MurNAc-L-Ala-D-Glu + L-Ala relative activity wild-type: 12% Mycobacterium tuberculosis ?
-
?
ATP + UDP-MurNAc-L-Ala-D-Glu + L-cystathionine relative activity wild-type: 12% Mycobacterium tuberculosis ?
-
?
ATP + UDP-MurNAc-L-Ala-D-Glu + L-Lys relative activity wild-type: 7% Mycobacterium tuberculosis ?
-
?
ATP + UDP-MurNAc-L-Ala-D-Glu + meso-2,6-diaminoheptanedioate relative activity wild-type: 100% Mycobacterium tuberculosis ADP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-D-gamma-glutamyl-meso-2,6-diaminoheptanedioate
-
?
GTP + UDP-MurNAc-L-Ala-D-Glu + meso-2,6-diaminoheptanedioate relative activity wild-type: 30% Mycobacterium tuberculosis GDP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-D-gamma-glutamyl-meso-2,6-diaminoheptanedioate
-
?
UTP + UDP-MurNAc-L-Ala-D-Glu + meso-2,6-diaminoheptanedioate relative activity wild-type: 17% Mycobacterium tuberculosis UDP + phosphate + UDP-N-acetylmuramoyl-L-alanyl-D-gamma-glutamyl-meso-2,6-diaminoheptanedioate
-
?

Synonyms

Synonyms Comment Organism
MurE
-
Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.05
-
ATP mutant N449D, pH 8.5, 37°C, co-substrate: UDP-MurNAc-L-Ala-D-Glu, meso-2,6-diaminoheptanedioate Mycobacterium tuberculosis
0.06
-
ATP mutant R451A, pH 8.5, 37°C, co-substrate: UDP-MurNAc-L-Ala-D-Glu, meso-2,6-diaminoheptanedioate Mycobacterium tuberculosis
0.08
-
ATP mutant K157A, pH 8.5, 37°C, co-substrate: UDP-MurNAc-L-Ala-D-Glu, meso-2,6-diaminoheptanedioate Mycobacterium tuberculosis
0.1
-
ATP mutant D392A, pH 8.5, 37°C, co-substrate: UDP-MurNAc-L-Ala-D-Glu, meso-2,6-diaminoheptanedioate Mycobacterium tuberculosis
0.56
-
ATP mutant E220A, pH 8.5, 37°C, co-substrate: UDP-MurNAc-L-Ala-D-Glu, meso-2,6-diaminoheptanedioate Mycobacterium tuberculosis
1.11
-
ATP N-terminal deletion mutant, pH 8.5, 37°C, co-substrate: UDP-MurNAc-L-Ala-D-Glu, meso-2,6-diaminoheptanedioate Mycobacterium tuberculosis
1.3
-
ATP wild-type, pH 8.5, 37°C, co-substrate: UDP-MurNAc-L-Ala-D-Glu, meso-2,6-diaminoheptanedioate Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
assay at Mycobacterium tuberculosis