3VGF
Crystal structure of glycosyltrehalose trehalohydrolase (D252S) complexed with maltotriosyltrehalose
3VGF の概要
エントリーDOI | 10.2210/pdb3vgf/pdb |
関連するPDBエントリー | 1EH9 3VGC 3VGD 3VGE 3VGG 3VGH |
関連するBIRD辞書のPRD_ID | PRD_900009 |
分子名称 | Malto-oligosyltrehalose trehalohydrolase, alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose, CITRATE ANION, ... (5 entities in total) |
機能のキーワード | alpha/beta barrel, trehalose, trehalohydrolase, alpha-amylase, hydrolase |
由来する生物種 | Sulfolobus solfataricus |
細胞内の位置 | Cytoplasm: Q55088 |
タンパク質・核酸の鎖数 | 1 |
化学式量合計 | 65959.82 |
構造登録者 | Okazaki, N.,Tamada, T.,Feese, M.D.,Kato, M.,Miura, Y.,Komeda, T.,Kobayashi, K.,Kondo, K.,Kuroki, R. (登録日: 2011-08-09, 公開日: 2012-06-20, 最終更新日: 2023-11-08) |
主引用文献 | Okazaki, N.,Tamada, T.,Feese, M.D.,Kato, M.,Miura, Y.,Komeda, T.,Kobayashi, K.,Kondo, K.,Blaber, M.,Kuroki, R. Substrate recognition mechanism of a glycosyltrehalose trehalohydrolase from Sulfolobus solfataricus KM1. Protein Sci., 21:539-552, 2012 Cited by PubMed Abstract: Glycosyltrehalose trehalohydrolase (GTHase) is an α-amylase that cleaves the α-1,4 bond adjacent to the α-1,1 bond of maltooligosyltrehalose to release trehalose. To investigate the catalytic and substrate recognition mechanisms of GTHase, two residues, Asp252 (nucleophile) and Glu283 (general acid/base), located at the catalytic site of GTHase were mutated (Asp252→Ser (D252S), Glu (D252E) and Glu283→Gln (E283Q)), and the activity and structure of the enzyme were investigated. The E283Q, D252E, and D252S mutants showed only 0.04, 0.03, and 0.6% of enzymatic activity against the wild-type, respectively. The crystal structure of the E283Q mutant GTHase in complex with the substrate, maltotriosyltrehalose (G3-Tre), was determined to 2.6-Å resolution. The structure with G3-Tre indicated that GTHase has at least five substrate binding subsites and that Glu283 is the catalytic acid, and Asp252 is the nucleophile that attacks the C1 carbon in the glycosidic linkage of G3-Tre. The complex structure also revealed a scheme for substrate recognition by GTHase. Substrate recognition involves two unique interactions: stacking of Tyr325 with the terminal glucose ring of the trehalose moiety and perpendicularly placement of Trp215 to the pyranose rings at the subsites -1 and +1 glucose. PubMed: 22334583DOI: 10.1002/pro.2039 主引用文献が同じPDBエントリー |
実験手法 | X-RAY DIFFRACTION (2.3 Å) |
構造検証レポート
検証レポート(詳細版)をダウンロード