Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D105A | site-directed mutagenesis, the mutant shows 50% reduced activity compared to the wild-type enzyme | Escherichia coli |
E154D | site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 80% reduced activity compared to the wild-type enzyme | Escherichia coli |
E154K | site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 90% reduced activity compared to the wild-type enzyme | Escherichia coli |
E154L | site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 70% reduced activity compared to the wild-type enzyme | Escherichia coli |
N169A | site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 50% reduced activity compared to the wild-type enzyme | Escherichia coli |
N169D | site-directed mutagenesis in the GlcNAc-binding region, the mutant shows unaltered activity compared to the wild-type enzyme | Escherichia coli |
N169Q | site-directed mutagenesis in the GlcNAc-binding region, the mutant shows unaltered activity compared to the wild-type enzyme | Escherichia coli |
N169R | site-directed mutagenesis in the GlcNAc-binding region, the mutant shows 1.4fold increased activity compared to the wild-type enzyme. The N169R mutant caused a slightly secondary structure changes, thus facilitating GlcNAc-1-phosphate to enter the active pocket through the additional interaction with N-acetyl arm of GlcNAc moiety | Escherichia coli |
Q76A | site-directed mutagenesis in the uridine-binding region, the mutant has a catalytic activity to convert CTP and GlcNAc-1P into unnatural sugar nucleotide CDP-GlcNAc which is distinct from the wild-type, altered nucleotide specificity compared to wild-type, overview | Escherichia coli |
Q76E | site-directed mutagenesis in the uridine-binding region, the mutant has a catalytic activity to convert CTP and GlcNAc-1P into unnatural sugar nucleotide CDP-GlcNAc which is distinct from the wild-type, altered nucleotide specificity compared to wild-type, overview | Escherichia coli |
Q76P | site-directed mutagenesis in the uridine-binding region, the mutant has a catalytic activity to convert CTP and GlcNAc-1P into unnatural sugar nucleotide CDP-GlcNAc which is distinct from the wild-type, altered nucleotide specificity compared to wild-type, overview | Escherichia coli |
T82G | site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 65% reduced activity compared to the wild-type enzyme | Escherichia coli |
T82Q | site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 80% reduced activity compared to the wild-type enzyme | Escherichia coli |
T82S | site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 80% reduced activity compared to the wild-type enzyme | Escherichia coli |
Y103F | site-directed mutagenesis of the residue located nearby the uridyltransferase active pocket,the mutant shows increased activity compared to the wild-type enzyme | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UTP + N-acetyl-alpha-D-glucosamine 1-phosphate | Escherichia coli | - |
diphosphate + UDP-N-acetyl-alpha-D-glucosamine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0ACC7 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
CTP + N-acetyl-alpha-D-glucosamine 1-phosphate | - |
Escherichia coli | diphosphate + CDP-N-acetyl-alpha-D-glucosamine | - |
? | |
additional information | the uridyltransferase domain of GlmU exhibits a flexible substrate specificity, roles of several highly conserved amino acid residues involved in substrate binding and recognition, overview. Besides UTP, the enzyme also shows activity with CTP, ATP, and slightly with dATP | Escherichia coli | ? | - |
? | |
UTP + N-acetyl-alpha-D-glucosamine 1-phosphate | - |
Escherichia coli | diphosphate + UDP-N-acetyl-alpha-D-glucosamine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
GlmU | - |
Escherichia coli |
N-acetylglucosamine-1-phosphate uridyltransferase | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |
General Information | Comment | Organism |
---|---|---|
metabolism | N-acetylglucosamine-1-phosphate uridyltransferase is a bifunctional enzyme that catalyzes both acetyltransfer and uridyltransfer reactions in the prokaryotic UDP-GlcNAc biosynthesis pathway | Escherichia coli |
additional information | the Tyr103-Asp105 segment is located at the floor of the uridyltransferase active pocket and is involved in interactions with UTP and GlcNAc-1-P substrates | Escherichia coli |
physiological function | N-acetylglucosamine-1-phosphate uridyltransferase (GlmU) is a bifunctional enzyme, its N- and C-terminal domains catalyze uridyltransferase, EC 2.7.7.23, and acetyltransferase, EC 2.3.1.157, activities, respectively. Final product of GlmU catalyzed reaction | Escherichia coli |