5Z66

Structure of periplasmic trehalase from Diamondback moth gut bacteria complexed with validoxylamine

5Z66 の概要

• エントリーDOI: 10.2210/pdb5z66/pdb
• 分子名称: Periplasmic trehalase, SULFATE ION, (1S,2S,3R,6S)-4-(HYDROXYMETHYL)-6-{[(1S,2S,3S,4R,5R)-2,3,4-TRIHYDROXY-5-(HYDROXYMETHYL)CYCLOHEXYL]AMINO}CYCLOHEX-4-ENE-1,2,3-TRIOL, ... (5 entities in total)
• 機能のキーワード: trehalase from insect gut bacterium, inhibitor validoxylamine-bound structure, hydrolase
• 由来する生物種: Enterobacter cloacae
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 64363.47

構造登録者

Harne, S.R.,Adhav, A.S.,Joshi, R.S.,Gayathri, P. (登録日: 2018-01-22, 公開日: 2019-01-23, 最終更新日: 2024-10-16)

主引用文献

Adhav, A.,Harne, S.,Bhide, A.,Giri, A.,Gayathri, P.,Joshi, R.
Mechanistic insights into enzymatic catalysis by trehalase from the insect gut endosymbiont Enterobacter cloacae.
Febs J., 286:1700-1716, 2019

PubMed Abstract: Energy metabolism in the diamondback moth Plutella xylostella is facilitated by trehalase, an enzyme which assists in trehalose hydrolysis, from the predominant gut bacterium Enterobacter cloacae. We report the biochemical and structural characterization of recombinant trehalase from E. cloacae (Px_EclTre). Px_EclTre showed K of 1.47 (±0.05) mm, k of 6254.72 min and V 0.2 (±0.002) mm·min at 55 °C and acidic pH. Crystal structures of Px_EclTre were determined in the ligand-free form and bound to the inhibitor Validoxylamine A. The crystal structure of the ligand-free form, unavailable until now for any other bacterial trehalases, enabled us to delineate the conformational changes accompanying ligand binding in trehalases. Multiple salt bridges were identified that potentially facilitated closure of a hood over the substrate-binding site. A cluster of five tryptophans lined the -1 substrate-binding subsite, interacted with crucial active site residues and contributed to both trehalase activity and stability. The importance of these residues in enzyme activity was further validated by mutagenesis studies. Many of these identified residues form part of signature motifs and other conserved sequences in trehalases. The structure analysis thus led to the assignment of the functional role to these conserved residues. This information can be further explored for the design of effective inhibitors against trehalases.

PubMed: 30657252
DOI: 10.1111/febs.14760

実験手法

X-RAY DIFFRACTION (1.8 Å)