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Literature summary extracted from

  • Woo, E.J.; Ryu, S.I.; Song, H.N.; Jung, T.Y.; Yeon, S.M.; Lee, H.A.; Park, B.C.; Park, K.H.; Lee, S.B.
    Structural insights on the new mechanism of trehalose synthesis by trehalose synthase TreT from Pyrococcus horikoshii (2010), J. Mol. Biol., 404, 247-259.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.245 gene treT, DNA and amino acid sequence determination and analysis, sequence comparison Pyrococcus horikoshii

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.1.245 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

EC Number Protein Variants Comment Organism
2.4.1.245 D134E the mutant shows 10% reduced activity in the forward reaction, and 50% reduced activity in the reverse reaction Pyrococcus horikoshii
2.4.1.245 D134R inactive mutant Pyrococcus horikoshii
2.4.1.245 D274R/D275A inactive mutant Pyrococcus horikoshii
2.4.1.245 E326A inactive mutant Pyrococcus horikoshii
2.4.1.245 F85R the mutant shows 95% reduced activity in the forward reaction, and no activity in the reverse reaction Pyrococcus horikoshii
2.4.1.245 F85Y the mutant shows 10% reduced activity in the forward reaction, and 40% reduced activity in the reverse reaction Pyrococcus horikoshii
2.4.1.245 H155D inactive in the forward reaction, 80% reduced activity in the reverse reaction compared to wild-type Pyrococcus horikoshii
2.4.1.245 H92A the mutant shows wild-type activity in the forward reaction, and 90% reduced activity in the reverse reaction Pyrococcus horikoshii
2.4.1.245 K209R the mutant shows wild-type activity Pyrococcus horikoshii
2.4.1.245 Q96A the mutant shows 10% reduced activity in the forward reaction, and 90% reduced activity in the reverse reaction Pyrococcus horikoshii
2.4.1.245 R239A inactive in the reverse reaction, 10% reduced activity in the forward reaction compared to wild-type Pyrococcus horikoshii
2.4.1.245 T49H the mutant shows 90% reduced activity in the forward reaction, and 98% reduced activity in the reverse reaction Pyrococcus horikoshii
2.4.1.245 T49R inactive mutant Pyrococcus horikoshii
2.4.1.245 V309L the mutant shows wild-type activity in the forward reaction, and 25% reduced activity in the reverse reaction Pyrococcus horikoshii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.245 ADP-alpha-D-glucose + D-glucose Pyrococcus horikoshii
-
 ADP + alpha,alpha-trehalose
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.245 Pyrococcus horikoshii O58762 gene treT
-

Reaction

EC Number Reaction Comment Organism Reaction ID
2.4.1.245 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
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.245 ADP-alpha-D-glucose + D-glucose
-
 Pyrococcus horikoshii ADP + alpha,alpha-trehalose
-
 r
2.4.1.245 GDP-glucose + D-glucose
-
 Pyrococcus horikoshii alpha,alpha-trehalose + GDP
-
 r
2.4.1.245 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 ?
-
 ?
2.4.1.245 UDP-glucose + D-glucose
-
 Pyrococcus horikoshii alpha,alpha-trehalose + UDP
-
 r

Synonyms

EC Number Synonyms Comment Organism
2.4.1.245 trehalose glycosyltransferring synthase
-
 Pyrococcus horikoshii
2.4.1.245 Trehalose synthase
-
 Pyrococcus horikoshii
2.4.1.245 TreT
-
 Pyrococcus horikoshii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.245 37
-
 assay at Pyrococcus horikoshii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.245 6
-
 assay at Pyrococcus horikoshii

General Information

EC Number General Information Comment Organism
2.4.1.245 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