2GDU

E232Q mutant of sucrose phosphorylase from BIFIDOBACTERIUM ADOLESCENTIS in complex with sucrose

2GDU の概要

• エントリーDOI: 10.2210/pdb2gdu/pdb
• 関連するPDBエントリー: 1R7A 2GDV
• 関連するBIRD辞書のPRD_ID: PRD_900003
• 分子名称: sucrose phosphorylase, beta-D-fructofuranose-(2-1)-alpha-D-glucopyranose (3 entities in total)
• 機能のキーワード: beta-alpha-barrels, dimer, glycoside hydrolase, transferase
• 由来する生物種: Bifidobacterium adolescentis
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 113251.96

構造登録者

Skov, L.K.,Mirza, O.,Gajhede, M.,Kastrup, J.S. (登録日: 2006-03-17, 公開日: 2006-09-26, 最終更新日: 2024-11-20)

主引用文献

Mirza, O.,Skov, L.K.,Sprogoe, D.,van den Broek, L.A.M.,Beldman, G.,Kastrup, J.S.,Gajhede, M.
Structural Rearrangements of Sucrose Phosphorylase from Bifidobacterium adolescentis during Sucrose Conversion
J.Biol.Chem., 281:35576-35584, 2006

PubMed Abstract: The reaction mechanism of sucrose phosphorylase from Bifidobacterium adolescentis (BiSP) was studied by site-directed mutagenesis and x-ray crystallography. An inactive mutant of BiSP (E232Q) was co-crystallized with sucrose. The structure revealed a substrate-binding mode comparable with that seen in other related sucrose-acting enzymes. Wild-type BiSP was also crystallized in the presence of sucrose. In the dimeric structure, a covalent glucosyl intermediate was formed in one molecule of the BiSP dimer, and after hydrolysis of the glucosyl intermediate, a beta-D-glucose product complex was formed in the other molecule. Although the overall structure of the BiSP-glucosyl intermediate complex is similar to that of the BiSP(E232Q)-sucrose complex, the glucose complex discloses major differences in loop conformations. Two loops (residues 336-344 and 132-137) in the proximity of the active site move up to 16 and 4 A, respectively. On the basis of these findings, we have suggested a reaction cycle that takes into account the large movements in the active-site entrance loops.

PubMed: 16990265
DOI: 10.1074/jbc.M605611200

実験手法

X-RAY DIFFRACTION (2.1 Å)