6TJQ

Crystal Structure of Recombinant GBA in Complex with 2-Deoxy-2-fluoro-beta-D-glucopyranoside

これはPDB形式変換不可エントリーです。

6TJQ の概要

• エントリーDOI: 10.2210/pdb6tjq/pdb
• 関連するPDBエントリー: 6TJJ 6TJK
• 分子名称: Glucosylceramidase, alpha-D-mannopyranose-(1-6)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, SULFATE ION, ... (7 entities in total)
• 機能のキーワード: beta-glucocerebrosidase, lysosomal glycoside hydrolase, gh30, hydrolase
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 58455.87

構造登録者

Rowland, R.J.,Davies, G.J. (登録日: 2019-11-26, 公開日: 2020-06-10, 最終更新日: 2024-11-20)

主引用文献

Rowland, R.J.,Wu, L.,Liu, F.,Davies, G.J.
A baculoviral system for the production of human beta-glucocerebrosidase enables atomic resolution analysis.
Acta Crystallogr D Struct Biol, 76:565-580, 2020

PubMed Abstract: The lysosomal glycoside hydrolase β-glucocerebrosidase (GBA; sometimes called GBA1 or GC) catalyses the hydrolysis of glycosphingolipids. Inherited deficiencies in GBA cause the lysosomal storage disorder Gaucher disease (GD). Consequently, GBA is of considerable medical interest, with continuous advances in the development of inhibitors, chaperones and activity-based probes. The development of new GBA inhibitors requires a source of active protein; however, the majority of structural and mechanistic studies of GBA today rely on clinical enzyme-replacement therapy (ERT) formulations, which are incredibly costly and are often difficult to obtain in adequate supply. Here, the production of active crystallizable GBA in insect cells using a baculovirus expression system is reported, providing a nonclinical source of recombinant GBA with comparable activity and biophysical properties to ERT preparations. Furthermore, a novel crystal form of GBA is described which diffracts to give a 0.98 Å resolution unliganded structure. A structure in complex with the inactivator 2,4-dinitrophenyl-2-deoxy-2-fluoro-β-D-glucopyranoside was also obtained, demonstrating the ability of this GBA formulation to be used in ligand-binding studies. In light of its purity, stability and activity, the GBA production protocol described here should circumvent the need for ERT formulations for structural and biochemical studies and serve to support GD research.

PubMed: 32496218
DOI: 10.1107/S205979832000501X

実験手法

X-RAY DIFFRACTION (1.41 Å)