Synthesis of modified 1,5-imino-d-xylitols as ligands for lysosomal β-glucocerebrosidase

Zoidl, Manuel; Wolfsgruber, Andreas; Schalli, Michael; Nasseri, Seyed A.; Weber, Patrick; Stütz, Arnold; Withers, Stephen G.; Wrodnigg, Tanja

doi:10.1007/s00706-019-02427-1

Cited by 5 publications

(5 citation statements)

References 38 publications

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“…These ERT preparations can be incredibly costly, and are often only obtainable under a Material Transfer Agreement (MTA) and in limited supply. Given the clinical importance of GBA and the continuing development of novel GBA chaperones (Goddard-Borger et al, 2012;Diot et al, 2011;Hill et al, 2011;Marugan et al, 2011), inhibitors (Artola et al, 2019;Kuo et al, 2019;Zoidl et al, 2019;Schrö der et al, 2018) and activity-based probes (ABPs; Artola et al, 2017Artola et al, , 2019Schrö der et al, 2018;Beenakker et al, 2017), there is a pressing need for reliable sources of recombinant GBA to meet research demands and reduce the reliance upon ERT formulations.…”

Section: Introductionmentioning

confidence: 99%

A baculoviral system for the production of human β-glucocerebrosidase enables atomic resolution analysis

Rowland¹,

Wu²,

Liu³

et al. 2020

Acta Crystallogr D Struct Biol

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The lysosomal glycoside hydrolase -glucocerebrosidase (GBA; sometimes called GBA1 or GC ase ) 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--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.

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Section: Introductionmentioning

confidence: 99%

A baculoviral system for the production of human β-glucocerebrosidase enables atomic resolution analysis

Rowland¹,

Wu²,

Liu³

et al. 2020

Acta Crystallogr D Struct Biol

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“…To investigate the importance of the enzyme specificity of the ligand we also synthesized and tested LDSE probes 5 and 6 with 1,5‐dideoxy‐1,5‐imino‐ d ‐xylitol (DIX, 16 ) as ligand (A). DIX 16 is another competitive inhibitor for β‐glucosidases [34–36] . DIX‐LDSE probes 5 and 6 showed lower affinity towards both model enzymes (Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…DIX 16 is another competitive inhibitor for β-glucosidases. [34][35][36] DIX-LDSE probes 5 and 6 showed lower affinity towards both model enzymes (Table 1). DIX-C 11 À LDSE-N 3 probe 6 forms a fluorescent covalent complex with PdGH1 after incubating for 20 h at 100 μM.…”

Section: Biologic Evaluation -Proof Of Conceptmentioning

confidence: 99%

Ligand‐Directed Chemistry on Glycoside Hydrolases – A Proof of Concept Study

Prasch,

Wolfsgruber,

Thonhofer

et al. 2023

ChemBioChem

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Selective covalent labelling of enzymes using small molecule probes has advanced the scopes of protein profiling. The covalent bond formation to a specific target is the key step of activity‐based protein profiling (ABPP), a method which has become an indispensable tool for measuring enzyme activity in complex matrices. With respect to carbohydrate processing enzymes, strategies for ABPP so far involve labelling the active site of the enzyme, which results in permanent loss of activity. Here, we report in a proof of concept study the use of ligand‐directed chemistry (LDC) for labelling glycoside hydrolases near to – but not in – the active site. During the labelling process, the competitive inhibitor is cleaved from the probe, departs the active site and the enzyme maintains its catalytic activity. To this end, we designed a building block synthetic concept for small molecule probes containing iminosugar‐based reversible inhibitors for labelling of two model β‐glucosidases. The results indicate that the LDC approach can be adaptable for covalent proximity labelling of glycoside hydrolases.

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“…In this context, a large variety of iminosugar based glycomimetics have been investigated [ 4 , 32 , 33 , 34 , 35 , 36 , 37 ]. Paradigmatic examples are shown in Figure 2 .…”

Section: Introductionmentioning

confidence: 99%

“…1,5- D ideoxy-1,5-imino- d -xylitol-based N -alkylated derivatives 19 and 20 were found as selective inhibitors of GCase with K i values of 57 µM and 4.1 µM, respectively. [ 36 ].…”

Section: Introductionmentioning

confidence: 99%

N-Alkylated Iminosugar Based Ligands: Synthesis and Inhibition of Human Lysosomal β-Glucocerebrosidase

et al. 2020

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The scope of a series of N-alkylated iminosugar based inhibitors in the d-gluco as well as d-xylo configuration towards their interaction with human lysosomal β-glucocerebrosidase has been evaluated. A versatile synthetic toolbox has been developed for the synthesis of N-alkylated iminosugar scaffolds conjugated to a variety of terminal groups via a benzoic acid ester linker. The terminal groups such as nitrile, azide, alkyne, nonafluoro-tert-butyl and amino substituents enable follow-up chemistry as well as visualisation experiments. All compounds showed promising inhibitory properties as well as selectivities for β-glucosidases, some exhibiting activities in the low nanomolar range for β-glucocerebrosidase.

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Synthesis of modified 1,5-imino-d-xylitols as ligands for lysosomal β-glucocerebrosidase

Cited by 5 publications

References 38 publications

A baculoviral system for the production of human β-glucocerebrosidase enables atomic resolution analysis

A baculoviral system for the production of human β-glucocerebrosidase enables atomic resolution analysis

Ligand‐Directed Chemistry on Glycoside Hydrolases – A Proof of Concept Study

N-Alkylated Iminosugar Based Ligands: Synthesis and Inhibition of Human Lysosomal β-Glucocerebrosidase

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