4WF7

Crystal structures of trehalose synthase from Deinococcus radiodurans reveal that a closed conformation is involved in the intramolecular isomerization catalysis

4WF7 の概要

• エントリーDOI: 10.2210/pdb4wf7/pdb
• 関連するPDBエントリー: 4TVU
• 分子名称: Trehalose synthase, CALCIUM ION, MAGNESIUM ION, ... (5 entities in total)
• 機能のキーワード: isomerase, trehalose synthase, glycoside hydrolase family 13, tris complex
• 由来する生物種: Deinococcus radiodurans R1
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 260647.53

構造登録者

Wang, Y.L.,Chow, S.Y.,Lin, Y.T.,Hsieh, Y.C.,Lee, G.C.,Liaw, S.H. (登録日: 2014-09-13, 公開日: 2014-12-24, 最終更新日: 2023-11-08)

主引用文献

Wang, Y.L.,Chow, S.Y.,Lin, Y.T.,Hsieh, Y.C.,Lee, G.C.,Liaw, S.H.
Structures of trehalose synthase from Deinococcus radiodurans reveal that a closed conformation is involved in catalysis of the intramolecular isomerization.
Acta Crystallogr.,Sect.D, 70:3144-3154, 2014

PubMed Abstract: Trehalose synthase catalyzes the simple conversion of the inexpensive maltose into trehalose with a side reaction of hydrolysis. Here, the crystal structures of the wild type and the N253A mutant of Deinococcus radiodurans trehalose synthase (DrTS) in complex with the inhibitor Tris are reported. DrTS consists of a catalytic (β/α)8 barrel, subdomain B, a C-terminal β domain and two TS-unique subdomains (S7 and S8). The C-terminal domain and S8 contribute the majority of the dimeric interface. DrTS shares high structural homology with sucrose hydrolase, amylosucrase and sucrose isomerase in complex with sucrose, in particular a virtually identical active-site architecture and a similar substrate-induced rotation of subdomain B. The inhibitor Tris was bound and mimics a sugar at the -1 subsite. A maltose was modelled into the active site, and subsequent mutational analysis suggested that Tyr213, Glu320 and Glu324 are essential within the +1 subsite for the TS activity. In addition, the interaction networks between subdomains B and S7 seal the active-site entrance. Disruption of such networks through the replacement of Arg148 and Asn253 with alanine resulted in a decrease in isomerase activity by 8-9-fold and an increased hydrolase activity by 1.5-1.8-fold. The N253A structure showed a small pore created for water entry. Therefore, our DrTS-Tris may represent a substrate-induced closed conformation that will facilitate intramolecular isomerization and minimize disaccharide hydrolysis.

PubMed: 25478833
DOI: 10.1107/S1399004714022500

実験手法

X-RAY DIFFRACTION (2.21 Å)