Cloned (Comment) | Organism |
---|---|
gene treT, DNA and amino acid sequence determination and analysis, sequence comparison | Pyrococcus horikoshii |
Crystallization (Comment) | Organism |
---|---|
purified enzyme free or as TreT-UDP binary complex, 10 mg/ml native enzyme from PEG 3350 25%, 0.2 M MgCl2, and 0.1 M sodium HEPES, 18°C, the selenomethionine-substituted protein crystal grow from 21% methoxy PEG 2000, 0.18 M ammonium sulfate, and 0.1 M sodium acetate, pH 4.6, for the UDP-glucose complex crystal, 5 mM UDPG is added to 10 mg/ml E326A protein for 1 h prior to setup of the crystallization using the same conditions as for the native crystal, X-ray diffraction structure determination and analysis at 2.3-3.0 A resolution, single-wavelength anomalous dispersion phasing | Pyrococcus horikoshii |
Protein Variants | Comment | Organism |
---|---|---|
D134E | the mutant shows 10% reduced activity in the forward reaction, and 50% reduced activity in the reverse reaction | Pyrococcus horikoshii |
D134R | inactive mutant | Pyrococcus horikoshii |
D274R/D275A | inactive mutant | Pyrococcus horikoshii |
E326A | inactive mutant | Pyrococcus horikoshii |
F85R | the mutant shows 95% reduced activity in the forward reaction, and no activity in the reverse reaction | Pyrococcus horikoshii |
F85Y | the mutant shows 10% reduced activity in the forward reaction, and 40% reduced activity in the reverse reaction | Pyrococcus horikoshii |
H155D | inactive in the forward reaction, 80% reduced activity in the reverse reaction compared to wild-type | Pyrococcus horikoshii |
H92A | the mutant shows wild-type activity in the forward reaction, and 90% reduced activity in the reverse reaction | Pyrococcus horikoshii |
K209R | the mutant shows wild-type activity | Pyrococcus horikoshii |
Q96A | the mutant shows 10% reduced activity in the forward reaction, and 90% reduced activity in the reverse reaction | Pyrococcus horikoshii |
R239A | inactive in the reverse reaction, 10% reduced activity in the forward reaction compared to wild-type | Pyrococcus horikoshii |
T49H | the mutant shows 90% reduced activity in the forward reaction, and 98% reduced activity in the reverse reaction | Pyrococcus horikoshii |
T49R | inactive mutant | Pyrococcus horikoshii |
V309L | the mutant shows wild-type activity in the forward reaction, and 25% reduced activity in the reverse reaction | Pyrococcus horikoshii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP-alpha-D-glucose + D-glucose | Pyrococcus horikoshii | - |
ADP + alpha,alpha-trehalose | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus horikoshii | O58762 | gene treT | - |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
NDP-alpha-D-glucose + D-glucose = alpha,alpha-trehalose + NDP | reaction mechanism, overview. The acceptor binding site of TreT shows a wide and commodious groove and lacks the long flexible loop that plays a gating role in ligand binding in trehalose phosphate synthase, TPS, EC 2.4.1.15. A wide space at the fissure between two domains and the relative shift of the N-domain in one of the crystal forms suggest that an interactive conformational change between two domains would occur, allowing a more compact architecture for catalysis | Pyrococcus horikoshii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP-alpha-D-glucose + D-glucose | - |
Pyrococcus horikoshii | ADP + alpha,alpha-trehalose | - |
r | |
GDP-glucose + D-glucose | - |
Pyrococcus horikoshii | alpha,alpha-trehalose + GDP | - |
r | |
additional information | molecular basis of the synthetic mechanism of trehalose, or the nucleotide sugar in the reverse reaction of TreT. TreT can utilize ADP-glucose and GDP-glucose as well as UDP-glucose to synthesize trehalose, the nucleotide sugar molecule is recognized by the nucleotide-sensing Gly-Gly-Leu motif that is located at the domain interface in many GT-B family members, and TreT contains this motif at residues 52-55, the acceptor molecule binds predominantly to the N-terminal domain | Pyrococcus horikoshii | ? | - |
? | |
UDP-glucose + D-glucose | - |
Pyrococcus horikoshii | alpha,alpha-trehalose + UDP | - |
r |
Synonyms | Comment | Organism |
---|---|---|
trehalose glycosyltransferring synthase | - |
Pyrococcus horikoshii |
Trehalose synthase | - |
Pyrococcus horikoshii |
TreT | - |
Pyrococcus horikoshii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Pyrococcus horikoshii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at | Pyrococcus horikoshii |
General Information | Comment | Organism |
---|---|---|
additional information | the acceptor binding site of TreT shows a wide and commodious groove and lacks the long flexible loop that plays a gating role in ligand binding in trehalose phosphate synthase, TPS, active site, and donor and acceptor binding pocket structure, overview. A wide space at the fissure between two domains and the relative shift of the N-domain in one of the crystal forms suggest that an interactive conformational change between two domains would occur, allowing a more compact architecture for catalysis | Pyrococcus horikoshii |