BRENDA - Enzyme Database
show all sequences of 2.3.1.157

Substrate bound crystal structures reveal features unique to Mycobacterium tuberculosis N-acetyl-glucosamine-1-phosphate uridyltransferase and a catalytic mechanism for acetyltransfer

Jagtap, P.K.; Soni, V.; Vithani, N.; Jhingan, G.D.; Bais, V.S.; Nandicoori, V.K.; Prakash, B.; J. Biol. Chem. 287, 39524-39537 (2012)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
additional information
the enzyme is regulated by PknB via phosphorylation at Thr418 causing downregulation of acetyltransferase
Mycobacterium tuberculosis
Application
Application
Commentary
Organism
drug development
the enzyme GlmU is a target for development of antibacterial drugs; the enzyme is a target for inhibitor development in treatment of tuberculosis
Mycobacterium tuberculosis
Crystallization (Commentary)
Crystallization
Organism
GlmUMtb in complex with substrates/products bound at the acetyltransferase active site, sitting drop vapor diffusion method, mixing of 400 nl of 15 mg/ml protein, 5 mM acetyl-CoA, 5 mM MgCl2, 5 mM UDP-GlcNAc with 400 nl of 18% PEG 3350, 0.1 M Tris-Cl, pH 8.5, and 2% tacsimate, 4-8 days, for enzyme complex with CoA and N-acetylglucosamine-1-phosphate, acetyl-Coa-containing crystals are soaked in 5 mM GlcN-1-P, 5 mM MgCl2, 5 mM UDP-GlcNAc, 5 mM acetyl-CoA, 18% PEG 3350, 0.1 M Tris-Cl, pH 8.5, and 2% tacsimate, or by co-crystallizing the enzyme with 5 mM GlcNAc-1-P, 5 mM MgCl2, 5 mM UDPGlcNAc, and 5 mM CoA under the conditions mentioned for obtaining GlmUMtb(AcCoA) crystals, X-ray diffraction structure determination and analysis at 1.98-2.33 A resolution; purified GlmU, sitting drop vapor diffusion method, mixing of 400 nl of 15 mg/ml GlmU in 5 mM acetyl-CoA, 5 mM MgCl2, 5 mM UDP-GlcNAc with 400 nL of 18% PEG 3350, 0.1 M Tris-Cl, pH 8.5, and 2% tacsimate, 7-8 days, for coupling to acetyl-CoA, crystals are soaked in 5 mM GlcN-1-P, 5 mM MgCl2, 5 mM UDP-GlcNAc, 5 mM acetyl-CoA, 18% PEG 3350, 0.1 M Tris-Cl, pH 8.5, and 2% tacsimate, or in 5 mM GlcNAc-1-P, 5 mM MgCl2, 5 mM UDP-GlcNAc, 5 mM CoA, X-ray diffraction structure determination and analysis at 1.98-2.33 A resolution
Mycobacterium tuberculosis
Engineering
Amino acid exchange
Commentary
Organism
A451R
site-directed mutagenesis, neither the single mutants A451R and R439T nor the double mutant A451R/R439T affect the acetyltransferase activity significantly
Mycobacterium tuberculosis
A451R/R439T
site-directed mutagenesis, neither the single mutants A451R and R439T nor the double mutant A451R/R439T affect the acetyltransferase activity significantly
Mycobacterium tuberculosis
H374A
site-directed mutagenesis, the acetyltransferase active site mutant shows 1.7% of acetyltransferase activity and 96.7% of uridinyltransferase activity compared to the wild-type; site-directed mutagenesis, the mutant shows highly reduced Vmax in acetyltransfer compared to the wild-type enzyme
Mycobacterium tuberculosis
K464A
site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme; site-directed mutagenesis, the mutant still shows acetyltransferase activity, the mutant shows 105.6% acetyltransferase activity and 97.9% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
K464A/W460A
site-directed mutagenesis, the mutant shows highly compromised activity compared to the wild-type enzyme
Mycobacterium tuberculosis
N397A
site-directed mutagenesis, the acetyltransferase active site mutant shows 5.2% of acetyltransferase activity and 113.6% of uridinyltransferase activity compared to the wild-type; site-directed mutagenesis, the mutant shows highly reduced Vmax in acetyltransfer compared to the wild-type enzyme
Mycobacterium tuberculosis
R439T
site-directed mutagenesis, neither the single mutants A451R and R439T nor the double mutant A451R/R439T affect the acetyltransferase activity significantly
Mycobacterium tuberculosis
S416A
site-directed mutagenesis, the acetyltransferase active site mutant shows 100.9% of acetyltransferase activity and 96.4% of uridinyltransferase activity compared to the wild-type; site-directed mutagenesis, the mutant shows kinetics in acetyltransfer similar to the wild-type enzyme, S416 neither plays a role in catalysis nor in substrate binding
Mycobacterium tuberculosis
T418A
site-directed mutagenesis, T418 is the most abundant phosphorylation site on GlmUMtb, acetyltransferase activity is completely abolishe; site-directed mutagenesis, the acetyltransferase activity of mutant is severely compromised as compared with GlmUMtb wild-type, the mutant shows 2.4% acetyltransferase activity and 100.4% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
T418E
site-directed mutagenesis, acetyltransferase activity of T418E mutant that mimics a phosphorylated Thr, is severely compromised compared to wild-type GlmUMtb; site-directed mutagenesis, the acetyltransferase activity of the T418E mutant that mimics a phosphorylated Thr, is severely compromised as compared with GlmUMtb wild-type, the mutant shows 2.2% acetyltransferase activity and 109.2% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
T418S
site-directed mutagenesis, the acetyltransferase activity of the mutant is compromised as compared with GlmUMtb wild-type, the mutant shows 19% acetyltransferase activity and 108.8% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
W460A
site-directed mutagenesis, the mutant displays almost complete loss in acetyltransferase activity, the mutant shows 8.4% acetyltransferase activity and 99.8% of uridinyltransferase activity compared to the wild-type; site-directed mutagenesis, the mutant shows highly compromised activity compared to the wild-type enzyme
Mycobacterium tuberculosis
W460A/K64A
site-directed mutagenesis, the mutant shows 7.8% acetyltransferase activity and 104.7% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.283
-
alpha-D-glucosamine 1-phosphate
pH 7.6, 30°C, recombinant mutant N397A
Mycobacterium tuberculosis
0.3
-
acetyl-CoA
pH 7.6, 30°C, recombinant mutant H374A
Mycobacterium tuberculosis
0.3
-
alpha-D-glucosamine 1-phosphate
pH 7.6, 30°C, recombinant mutant H374A
Mycobacterium tuberculosis
0.353
-
alpha-D-glucosamine 1-phosphate
pH 7.6, 30°C, recombinant wild-type
Mycobacterium tuberculosis
0.355
-
acetyl-CoA
pH 7.6, 30°C, recombinant mutant N397A; pH 7.6, 30°C, recombinant mutant S416A; pH 7.6, 30°C, recombinant wild-type
Mycobacterium tuberculosis
0.37
-
alpha-D-glucosamine 1-phosphate
pH 7.6, 30°C, recombinant mutant S416A
Mycobacterium tuberculosis
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
acetyl-CoA + alpha-D-glucosamine 1-phosphate
Mycobacterium tuberculosis
-
CoA + N-acetyl-alpha-D-glucosamine 1-phosphate
-
-
?
additional information
Mycobacterium tuberculosis
N-acetyl-glucosamine-1-phosphate uridyltransferase, GlmU, a bifunctional enzyme that catalyzes two key reactions: acetyltransfer and uridyltransfer at two independent domains, overview
?
-
-
-
additional information
Mycobacterium tuberculosis
N-acetylglucosamine-1-phosphate uridyltransferase (GlmU) is a bifunctional enzyme catalyzing the reactions of EC 2.3.1.157, N-acetylglucosamine-1-phosphate uridyltransferase, and 2.7.7.23, UDP-N-acetylglucosamine diphosphorylase, the enzyme catalyzes the two reactions, acetyl transfer and uridyl transfer, at two independent domains, regulation, overview
?
-
-
-
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Mycobacterium tuberculosis
-
-
-
Mycobacterium tuberculosis
P9WMN3
-
-
Posttranslational Modification
Posttranslational Modification
Commentary
Organism
phosphoprotein
the enzyme is regulated by PknB via phosphorylation at Thr418 causing downregulation of acetyltransferase activity leaving its uridyltransferase activity unaffected, identification of phosphorylation site by mass spectrometry
Mycobacterium tuberculosis
Reaction
Reaction
Commentary
Organism
acetyl-CoA + alpha-D-glucosamine 1-phosphate = CoA + N-acetyl-alpha-D-glucosamine 1-phosphate
substrate recognition, and catalytic mechanism for acetyltransfer involving His374, Asn397 and Ala391, overview; the catalytic mechanism is a SN2, bimolecular nucleophilic substitution reaction, catalyzed by the C-terminal domain. His374 and Asn397 act as catalytic residues by enhancing the nucleophilicity of the attacking amino group of glucosamine 1-phosphate. Ser416 and Trp460, on a short helix, provide important interactions for substrate binding
Mycobacterium tuberculosis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
acetyl-CoA + alpha-D-glucosamine 1-phosphate
-
720042
Mycobacterium tuberculosis
CoA + N-acetyl-alpha-D-glucosamine 1-phosphate
-
-
-
?
acetyl-CoA + alpha-D-glucosamine 1-phosphate
uncommon mode of acetyl-CoA binding in GlmUMtb in the U conformation, which is distinct from the L conformation seen in the available non-mycobacterial GlmU structures. Higly conserved Trp460 is critical for acetyl-CoA binding
720042
Mycobacterium tuberculosis
CoA + N-acetyl-alpha-D-glucosamine 1-phosphate
-
-
-
?
additional information
N-acetyl-glucosamine-1-phosphate uridyltransferase, GlmU, a bifunctional enzyme that catalyzes two key reactions: acetyltransfer and uridyltransfer at two independent domains, overview
720042
Mycobacterium tuberculosis
?
-
-
-
-
additional information
N-acetylglucosamine-1-phosphate uridyltransferase (GlmU) is a bifunctional enzyme catalyzing the reactions of EC 2.3.1.157, N-acetylglucosamine-1-phosphate uridyltransferase, and 2.7.7.23, UDP-N-acetylglucosamine diphosphorylase, the enzyme catalyzes the two reactions, acetyl transfer and uridyl transfer, at two independent domains, regulation, overview
720042
Mycobacterium tuberculosis
?
-
-
-
-
additional information
coupled assay method: coupling of the two enzyme reactions via N-acetyl-alpha-D-glucosamine 1-phosphate for determination of the acetyl transferase activity of the enzyme. Substrate recognition and catalytic mechanism for acetyl transfer, overview
720042
Mycobacterium tuberculosis
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
trimer
GlmU has a conserved two-domain architecture, one monomer per asymmetric unit, and a trimeric quaternary structure, modeling of acetyl-CoA bound to the C-terminal domain, overview; two-domain architecture of GlmU, one monomer per asymmetric unit, and a trimeric quaternary structure known for GlmU proteins
Mycobacterium tuberculosis
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at; assay at
Mycobacterium tuberculosis
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.6
-
assay at; assay at
Mycobacterium tuberculosis
Cofactor
Cofactor
Commentary
Organism
Structure
acetyl-CoA
-
Mycobacterium tuberculosis
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
additional information
the enzyme is regulated by PknB via phosphorylation at Thr418 causing downregulation of acetyltransferase
Mycobacterium tuberculosis
Application (protein specific)
Application
Commentary
Organism
drug development
the enzyme GlmU is a target for development of antibacterial drugs
Mycobacterium tuberculosis
drug development
the enzyme is a target for inhibitor development in treatment of tuberculosis
Mycobacterium tuberculosis
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
acetyl-CoA
-
Mycobacterium tuberculosis
Crystallization (Commentary) (protein specific)
Crystallization
Organism
GlmUMtb in complex with substrates/products bound at the acetyltransferase active site, sitting drop vapor diffusion method, mixing of 400 nl of 15 mg/ml protein, 5 mM acetyl-CoA, 5 mM MgCl2, 5 mM UDP-GlcNAc with 400 nl of 18% PEG 3350, 0.1 M Tris-Cl, pH 8.5, and 2% tacsimate, 4-8 days, for enzyme complex with CoA and N-acetylglucosamine-1-phosphate, acetyl-Coa-containing crystals are soaked in 5 mM GlcN-1-P, 5 mM MgCl2, 5 mM UDP-GlcNAc, 5 mM acetyl-CoA, 18% PEG 3350, 0.1 M Tris-Cl, pH 8.5, and 2% tacsimate, or by co-crystallizing the enzyme with 5 mM GlcNAc-1-P, 5 mM MgCl2, 5 mM UDPGlcNAc, and 5 mM CoA under the conditions mentioned for obtaining GlmUMtb(AcCoA) crystals, X-ray diffraction structure determination and analysis at 1.98-2.33 A resolution
Mycobacterium tuberculosis
purified GlmU, sitting drop vapor diffusion method, mixing of 400 nl of 15 mg/ml GlmU in 5 mM acetyl-CoA, 5 mM MgCl2, 5 mM UDP-GlcNAc with 400 nL of 18% PEG 3350, 0.1 M Tris-Cl, pH 8.5, and 2% tacsimate, 7-8 days, for coupling to acetyl-CoA, crystals are soaked in 5 mM GlcN-1-P, 5 mM MgCl2, 5 mM UDP-GlcNAc, 5 mM acetyl-CoA, 18% PEG 3350, 0.1 M Tris-Cl, pH 8.5, and 2% tacsimate, or in 5 mM GlcNAc-1-P, 5 mM MgCl2, 5 mM UDP-GlcNAc, 5 mM CoA, X-ray diffraction structure determination and analysis at 1.98-2.33 A resolution
Mycobacterium tuberculosis
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
A451R
site-directed mutagenesis, neither the single mutants A451R and R439T nor the double mutant A451R/R439T affect the acetyltransferase activity significantly
Mycobacterium tuberculosis
A451R/R439T
site-directed mutagenesis, neither the single mutants A451R and R439T nor the double mutant A451R/R439T affect the acetyltransferase activity significantly
Mycobacterium tuberculosis
H374A
site-directed mutagenesis, the acetyltransferase active site mutant shows 1.7% of acetyltransferase activity and 96.7% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
H374A
site-directed mutagenesis, the mutant shows highly reduced Vmax in acetyltransfer compared to the wild-type enzyme
Mycobacterium tuberculosis
K464A
site-directed mutagenesis, the mutant still shows acetyltransferase activity, the mutant shows 105.6% acetyltransferase activity and 97.9% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
K464A
site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme
Mycobacterium tuberculosis
K464A/W460A
site-directed mutagenesis, the mutant shows highly compromised activity compared to the wild-type enzyme
Mycobacterium tuberculosis
N397A
site-directed mutagenesis, the acetyltransferase active site mutant shows 5.2% of acetyltransferase activity and 113.6% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
N397A
site-directed mutagenesis, the mutant shows highly reduced Vmax in acetyltransfer compared to the wild-type enzyme
Mycobacterium tuberculosis
R439T
site-directed mutagenesis, neither the single mutants A451R and R439T nor the double mutant A451R/R439T affect the acetyltransferase activity significantly
Mycobacterium tuberculosis
S416A
site-directed mutagenesis, the acetyltransferase active site mutant shows 100.9% of acetyltransferase activity and 96.4% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
S416A
site-directed mutagenesis, the mutant shows kinetics in acetyltransfer similar to the wild-type enzyme, S416 neither plays a role in catalysis nor in substrate binding
Mycobacterium tuberculosis
T418A
site-directed mutagenesis, the acetyltransferase activity of mutant is severely compromised as compared with GlmUMtb wild-type, the mutant shows 2.4% acetyltransferase activity and 100.4% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
T418A
site-directed mutagenesis, T418 is the most abundant phosphorylation site on GlmUMtb, acetyltransferase activity is completely abolishe
Mycobacterium tuberculosis
T418E
site-directed mutagenesis, the acetyltransferase activity of the T418E mutant that mimics a phosphorylated Thr, is severely compromised as compared with GlmUMtb wild-type, the mutant shows 2.2% acetyltransferase activity and 109.2% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
T418E
site-directed mutagenesis, acetyltransferase activity of T418E mutant that mimics a phosphorylated Thr, is severely compromised compared to wild-type GlmUMtb
Mycobacterium tuberculosis
T418S
site-directed mutagenesis, the acetyltransferase activity of the mutant is compromised as compared with GlmUMtb wild-type, the mutant shows 19% acetyltransferase activity and 108.8% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
W460A
site-directed mutagenesis, the mutant displays almost complete loss in acetyltransferase activity, the mutant shows 8.4% acetyltransferase activity and 99.8% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
W460A
site-directed mutagenesis, the mutant shows highly compromised activity compared to the wild-type enzyme
Mycobacterium tuberculosis
W460A/K64A
site-directed mutagenesis, the mutant shows 7.8% acetyltransferase activity and 104.7% of uridinyltransferase activity compared to the wild-type
Mycobacterium tuberculosis
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.283
-
alpha-D-glucosamine 1-phosphate
pH 7.6, 30°C, recombinant mutant N397A
Mycobacterium tuberculosis
0.3
-
acetyl-CoA
pH 7.6, 30°C, recombinant mutant H374A
Mycobacterium tuberculosis
0.3
-
alpha-D-glucosamine 1-phosphate
pH 7.6, 30°C, recombinant mutant H374A
Mycobacterium tuberculosis
0.353
-
alpha-D-glucosamine 1-phosphate
pH 7.6, 30°C, recombinant wild-type
Mycobacterium tuberculosis
0.355
-
acetyl-CoA
pH 7.6, 30°C, recombinant mutant N397A; pH 7.6, 30°C, recombinant mutant S416A; pH 7.6, 30°C, recombinant wild-type
Mycobacterium tuberculosis
0.37
-
alpha-D-glucosamine 1-phosphate
pH 7.6, 30°C, recombinant mutant S416A
Mycobacterium tuberculosis
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
acetyl-CoA + alpha-D-glucosamine 1-phosphate
Mycobacterium tuberculosis
-
CoA + N-acetyl-alpha-D-glucosamine 1-phosphate
-
-
?
additional information
Mycobacterium tuberculosis
N-acetyl-glucosamine-1-phosphate uridyltransferase, GlmU, a bifunctional enzyme that catalyzes two key reactions: acetyltransfer and uridyltransfer at two independent domains, overview
?
-
-
-
additional information
Mycobacterium tuberculosis
N-acetylglucosamine-1-phosphate uridyltransferase (GlmU) is a bifunctional enzyme catalyzing the reactions of EC 2.3.1.157, N-acetylglucosamine-1-phosphate uridyltransferase, and 2.7.7.23, UDP-N-acetylglucosamine diphosphorylase, the enzyme catalyzes the two reactions, acetyl transfer and uridyl transfer, at two independent domains, regulation, overview
?
-
-
-
Posttranslational Modification (protein specific)
Posttranslational Modification
Commentary
Organism
phosphoprotein
the enzyme is regulated by PknB via phosphorylation at Thr418 causing downregulation of acetyltransferase activity leaving its uridyltransferase activity unaffected, identification of phosphorylation site by mass spectrometry
Mycobacterium tuberculosis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
acetyl-CoA + alpha-D-glucosamine 1-phosphate
-
720042
Mycobacterium tuberculosis
CoA + N-acetyl-alpha-D-glucosamine 1-phosphate
-
-
-
?
acetyl-CoA + alpha-D-glucosamine 1-phosphate
uncommon mode of acetyl-CoA binding in GlmUMtb in the U conformation, which is distinct from the L conformation seen in the available non-mycobacterial GlmU structures. Higly conserved Trp460 is critical for acetyl-CoA binding
720042
Mycobacterium tuberculosis
CoA + N-acetyl-alpha-D-glucosamine 1-phosphate
-
-
-
?
additional information
N-acetyl-glucosamine-1-phosphate uridyltransferase, GlmU, a bifunctional enzyme that catalyzes two key reactions: acetyltransfer and uridyltransfer at two independent domains, overview
720042
Mycobacterium tuberculosis
?
-
-
-
-
additional information
N-acetylglucosamine-1-phosphate uridyltransferase (GlmU) is a bifunctional enzyme catalyzing the reactions of EC 2.3.1.157, N-acetylglucosamine-1-phosphate uridyltransferase, and 2.7.7.23, UDP-N-acetylglucosamine diphosphorylase, the enzyme catalyzes the two reactions, acetyl transfer and uridyl transfer, at two independent domains, regulation, overview
720042
Mycobacterium tuberculosis
?
-
-
-
-
additional information
coupled assay method: coupling of the two enzyme reactions via N-acetyl-alpha-D-glucosamine 1-phosphate for determination of the acetyl transferase activity of the enzyme. Substrate recognition and catalytic mechanism for acetyl transfer, overview
720042
Mycobacterium tuberculosis
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
trimer
two-domain architecture of GlmU, one monomer per asymmetric unit, and a trimeric quaternary structure known for GlmU proteins
Mycobacterium tuberculosis
trimer
GlmU has a conserved two-domain architecture, one monomer per asymmetric unit, and a trimeric quaternary structure, modeling of acetyl-CoA bound to the C-terminal domain, overview
Mycobacterium tuberculosis
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Mycobacterium tuberculosis
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.6
-
assay at
Mycobacterium tuberculosis
General Information
General Information
Commentary
Organism
evolution
N-acetyl-glucosamine-1-phosphate uridyltransferase, GlmU, is exclusive to prokaryotes, conserved both in Gram positive and Gram negative bacteria
Mycobacterium tuberculosis
malfunction
Deleting the C-terminal tail, i.e. residues 457-495, of GlmUMtb that provides these residues abolishes all acetyltransferase activity
Mycobacterium tuberculosis
metabolism
N-acetyl-glucosamine-1-phosphate uridyltransferase, GlmU, is a bifunctional enzyme involved in bacterial cell wall synthesis
Mycobacterium tuberculosis
additional information
analysis of structures of GlmUMtb bound to substrates of the acetyl transfer reaction; the catalytic mechanism operative in GlmUMtb performs a SN2 reaction, His374 and Asn397 act as catalytic residues by enhancing the nucleophilicity of the attacking amino group of glucosamine 1-phosphate. Ser416 and Trp460, on a short helix, provide important interactions for substrate binding. The enzyme shows an uncommon mode of binding with acetyl-CoA. GlmU from Mycobacterium tuberculosis possesses a unique 30-residue extension at the C-terminus. The adenine base of acetyl-CoA bound to GlmUMtb is buried at the interface of two monomers of the trimer
Mycobacterium tuberculosis
physiological function
GlmU is involved in the biosynthesis of UDP-N-acetylglucosamine-1-phosphate. It is a bifunctional protein with two independent active sites catalyzing acetyl transfer and uridyl transfer reactions on glucosamine-1-phosphate. It synthesizes two key intermediates of cell wall biosynthesis pathways, viz. N-acetylglucosamine-1-phosphate (GlcNAc-1-P) and UDP-GlcNAc. The acetyltransferase activity is catalyzed by the C-terminal domain. It is essential for the growth of the organism; N-acetyl-glucosamine-1-phosphate uridyltransferase (GlmU) is a bifunctional enzyme involved in bacterial cell wall synthesis and is exclusive to prokaryotes. The enzyme is regulated by PknB via phosphorylation at Thr418 causing downregulation of acetyltransferase
Mycobacterium tuberculosis
General Information (protein specific)
General Information
Commentary
Organism
evolution
N-acetyl-glucosamine-1-phosphate uridyltransferase, GlmU, is exclusive to prokaryotes, conserved both in Gram positive and Gram negative bacteria
Mycobacterium tuberculosis
malfunction
Deleting the C-terminal tail, i.e. residues 457-495, of GlmUMtb that provides these residues abolishes all acetyltransferase activity
Mycobacterium tuberculosis
metabolism
N-acetyl-glucosamine-1-phosphate uridyltransferase, GlmU, is a bifunctional enzyme involved in bacterial cell wall synthesis
Mycobacterium tuberculosis
additional information
the catalytic mechanism operative in GlmUMtb performs a SN2 reaction, His374 and Asn397 act as catalytic residues by enhancing the nucleophilicity of the attacking amino group of glucosamine 1-phosphate. Ser416 and Trp460, on a short helix, provide important interactions for substrate binding. The enzyme shows an uncommon mode of binding with acetyl-CoA. GlmU from Mycobacterium tuberculosis possesses a unique 30-residue extension at the C-terminus. The adenine base of acetyl-CoA bound to GlmUMtb is buried at the interface of two monomers of the trimer
Mycobacterium tuberculosis
additional information
analysis of structures of GlmUMtb bound to substrates of the acetyl transfer reaction
Mycobacterium tuberculosis
physiological function
N-acetyl-glucosamine-1-phosphate uridyltransferase (GlmU) is a bifunctional enzyme involved in bacterial cell wall synthesis and is exclusive to prokaryotes. The enzyme is regulated by PknB via phosphorylation at Thr418 causing downregulation of acetyltransferase
Mycobacterium tuberculosis
physiological function
GlmU is involved in the biosynthesis of UDP-N-acetylglucosamine-1-phosphate. It is a bifunctional protein with two independent active sites catalyzing acetyl transfer and uridyl transfer reactions on glucosamine-1-phosphate. It synthesizes two key intermediates of cell wall biosynthesis pathways, viz. N-acetylglucosamine-1-phosphate (GlcNAc-1-P) and UDP-GlcNAc. The acetyltransferase activity is catalyzed by the C-terminal domain. It is essential for the growth of the organism
Mycobacterium tuberculosis
Other publictions for EC 2.3.1.157
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
746844
Honda
Improvement of ST0452 N-acety ...
Sulfurisphaera tokodaii, Sulfurisphaera tokodaii DSM 16993
Appl. Environ. Microbiol.
84
e002213-18
2018
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10
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1
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1
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1
2
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1
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736517
Sharma
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7
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1
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9
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1
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6
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9
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11
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8
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1
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2
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2
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14
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1
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1
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Verma
Structure of N-acetylglucosami ...
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1
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