5LVX

Crystal structure of glucocerebrosidase with an inhibitory quinazoline modulator

5LVX の概要

• エントリーDOI: 10.2210/pdb5lvx/pdb
• 分子名称: Glucosylceramidase, alpha-D-mannopyranose-(1-2)-alpha-D-mannopyranose-(1-2)-alpha-D-mannopyranose-(1-3)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, ... (9 entities in total)
• 機能のキーワード: gcase, hydrolase, glycoprotein, glycosidase, glucosylceramidase, prot, proteros biostructures gmbh
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 230629.43

構造登録者

Zheng, J.,Chen, L.,Skinner, O.S.,Lansbury, P.,Skerlj, R.,Mrosek, M.,Heunisch, U.,Krapp, S.,Weigand, S.,Charrow, J.,Schwake, M.,Kelleher, N.L.,Silverman, R.B.,Krainc, D. (登録日: 2016-09-14, 公開日: 2017-10-25, 最終更新日: 2024-10-23)

主引用文献

Zheng, J.,Chen, L.,Skinner, O.S.,Ysselstein, D.,Remis, J.,Lansbury, P.,Skerlj, R.,Mrosek, M.,Heunisch, U.,Krapp, S.,Charrow, J.,Schwake, M.,Kelleher, N.L.,Silverman, R.B.,Krainc, D.
beta-Glucocerebrosidase Modulators Promote Dimerization of beta-Glucocerebrosidase and Reveal an Allosteric Binding Site.
J. Am. Chem. Soc., 140:5914-5924, 2018

PubMed Abstract: β-Glucocerebrosidase (GCase) mutations cause Gaucher's disease and are a high risk factor in Parkinson's disease. The implementation of a small molecule modulator is a strategy to restore proper folding and lysosome delivery of degradation-prone mutant GCase. Here, we present a potent quinazoline modulator, JZ-4109, which stabilizes wild-type and N370S mutant GCase and increases GCase abundance in patient-derived fibroblast cells. We then developed a covalent modification strategy using a lysine targeted inactivator (JZ-5029) for in vitro mechanistic studies. By using native top-down mass spectrometry, we located two potentially covalently modified lysines. We obtained the first crystal structure, at 2.2 Å resolution, of a GCase with a noniminosugar modulator covalently bound, and were able to identify the exact lysine residue modified (Lys346) and reveal an allosteric binding site. GCase dimerization was induced by our modulator binding, which was observed by native mass spectrometry, its crystal structure, and size exclusion chromatography with a multiangle light scattering detector. Finally, the dimer form was confirmed by negative staining transmission electron microscopy studies. Our newly discovered allosteric site and observed GCase dimerization provide a new mechanistic insight into GCase and its noniminosugar modulators and facilitate the rational design of novel GCase modulators for Gaucher's disease and Parkinson's disease.

PubMed: 29676907
DOI: 10.1021/jacs.7b13003

実験手法

X-RAY DIFFRACTION (2.2 Å)