Cloned (Comment) | Organism |
---|---|
gene thrB, recombinant expression of His-taged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Corynebacterium glutamicum |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged enzyme in apoform or with either methionine or ATPgammaS and cysteine, hanging drop vapor diffusion method, mixing of 0.002 ml of 16.2 mg/ml protein in 20 mM Tris, pH 7.5, 150 mM NaCl, 50 mM KCl, and 50 mM MgCl2 with 0.002 ml of well solution containing 0.25 M ammonium sulfate, 25% PEG 3350, and 0.1 M HEPES, pH 7.5, and equilibration against 0.1 ml of well solution, X-ray diffraction structure determination and analysis at 1.8-2.2 A resolution | Corynebacterium glutamicum |
Protein Variants | Comment | Organism |
---|---|---|
A20G | site-directed mutagenesis, the mutant retains wild-type enzymatic activity, with dramatically decreased feedback inhibition by L-threonine. The changes in L-threonine affinity to the CglThrB-A20G active site derive from loss of van der Waals interactions | Corynebacterium glutamicum |
A20L | site-directed mutagenesis | Corynebacterium glutamicum |
A20S | site-directed mutagenesis | Corynebacterium glutamicum |
A20V | site-directed mutagenesis | Corynebacterium glutamicum |
additional information | engineering of Corynebacterium glutamicum to increase production of L-threonine, the attempt is to render the active site of Corynebacterium glutamicum ThrB (CglThrB) more selective toward L-homoserine than L-threonine | Corynebacterium glutamicum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
L-threonine | competitive inhibition, interaction with ThrB residue A20 is important. Highly reduced inhibition of enzyme mutant A20G. The changes in L-threonine affinity to the CglThrB-A20G active site derive from loss of van der Waals interactions | Corynebacterium glutamicum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Corynebacterium glutamicum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-homoserine | Corynebacterium glutamicum | - |
ADP + O-phospho-L-homoserine | - |
? | |
ATP + L-homoserine | Corynebacterium glutamicum LMG 3730 | - |
ADP + O-phospho-L-homoserine | - |
? | |
ATP + L-homoserine | Corynebacterium glutamicum BCRC 11384 | - |
ADP + O-phospho-L-homoserine | - |
? | |
ATP + L-homoserine | Corynebacterium glutamicum ATCC 13032 | - |
ADP + O-phospho-L-homoserine | - |
? | |
ATP + L-homoserine | Corynebacterium glutamicum JCM 1318 | - |
ADP + O-phospho-L-homoserine | - |
? | |
ATP + L-homoserine | Corynebacterium glutamicum NCIMB 10025 | - |
ADP + O-phospho-L-homoserine | - |
? | |
ATP + L-homoserine | Corynebacterium glutamicum DSM 20300 | - |
ADP + O-phospho-L-homoserine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Corynebacterium glutamicum | P07128 | - |
- |
Corynebacterium glutamicum ATCC 13032 | P07128 | - |
- |
Corynebacterium glutamicum BCRC 11384 | P07128 | - |
- |
Corynebacterium glutamicum DSM 20300 | P07128 | - |
- |
Corynebacterium glutamicum JCM 1318 | P07128 | - |
- |
Corynebacterium glutamicum LMG 3730 | P07128 | - |
- |
Corynebacterium glutamicum NCIMB 10025 | P07128 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-taged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis | Corynebacterium glutamicum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-homoserine | - |
Corynebacterium glutamicum | ADP + O-phospho-L-homoserine | - |
? | |
ATP + L-homoserine | - |
Corynebacterium glutamicum LMG 3730 | ADP + O-phospho-L-homoserine | - |
? | |
ATP + L-homoserine | - |
Corynebacterium glutamicum BCRC 11384 | ADP + O-phospho-L-homoserine | - |
? | |
ATP + L-homoserine | - |
Corynebacterium glutamicum ATCC 13032 | ADP + O-phospho-L-homoserine | - |
? | |
ATP + L-homoserine | - |
Corynebacterium glutamicum JCM 1318 | ADP + O-phospho-L-homoserine | - |
? | |
ATP + L-homoserine | - |
Corynebacterium glutamicum NCIMB 10025 | ADP + O-phospho-L-homoserine | - |
? | |
ATP + L-homoserine | - |
Corynebacterium glutamicum DSM 20300 | ADP + O-phospho-L-homoserine | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | Corynebacterium glutamicum CbThrB crystal structure (PDB ID 5WAT) comparison with the crystal structure of Methanocaldococcus jannaschii ThrB with bound L-Thr or L-homoserine (MjaThrB, PDB ID 1H72 or 1H73), overview | Corynebacterium glutamicum |
Synonyms | Comment | Organism |
---|---|---|
CglThrB | - |
Corynebacterium glutamicum |
CThrB | - |
Corynebacterium glutamicum |
ThrB | - |
Corynebacterium glutamicum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
27 | - |
assay at | Corynebacterium glutamicum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.8 | - |
assay at | Corynebacterium glutamicum |
General Information | Comment | Organism |
---|---|---|
additional information | a conserved active-site alanine residue, A20, in CThrB (CglThrB) is important for differential interactions with L-threonine and L-homoserine | Corynebacterium glutamicum |