1,6-Cyclophellitol Cyclosulfates: A New Class of Irreversible Glycosidase Inhibitor

Artola, Marta; Wu, Liang; Ferraz, Maria J.; Kuo, Chi Lin; Raich, Lluís; Breen, Imogen; Offen, W.A.; Codée, Jeroen D. C.; Marel, Gijsbert A. van der; Rovira, Carme; Aerts, Johannes M. F. G.; Davies, G.J.; Overkleeft, Herman S.

doi:10.1021/acscentsci.7b00214

Cited by 44 publications

(90 citation statements)

References 40 publications

(88 reference statements)

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“…The conformations can strongly influence the inhibitory potencies and selectivities of cyclophellitol analogues . Inhibitors 1 – 3 are slower binders than the corresponding cyclophellitol aziridine conformers (such as β‐ gluco ‐aziridine ABP JJB367 for GBA).…”

Section: Discussionmentioning

confidence: 99%

New Irreversible α‐l‐Iduronidase Inhibitors and Activity‐Based Probes

Artola

Kuo

McMahon

et al. 2018

Chemistry A European J

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Cyclophellitol aziridines are potent irreversible inhibitors of retaining glycosidases and versatile intermediates in the synthesis of activity‐based glycosidase probes (ABPs). Direct 3‐amino‐2‐(trifluoromethyl)quinazolin‐4(3H)‐one‐mediated aziridination of l‐ido‐configured cyclohexene has enabled the synthesis of new covalent inhibitors and ABPs of α‐l‐iduronidase, deficiency of which underlies the lysosomal storage disorder mucopolysaccharidosis type I (MPS I). The iduronidase ABPs react covalently and irreversibly in an activity‐based manner with human recombinant α‐l‐iduronidase (rIDUA, Aldurazyme®). The structures of IDUA when complexed with the inhibitors in a non‐covalent transition state mimicking form and a covalent enzyme‐bound form provide insights into its conformational itinerary. Inhibitors 1–3 adopt a half‐chair conformation in solution (4H3 and 3H4), as predicted by DFT calculations, which is different from the conformation of the Michaelis complex observed by crystallographic studies. Consequently, 1–3 may need to overcome an energy barrier in order to switch from the 4H3 conformation to the transition state (2, 5B) binding conformation before reacting and adopting a covalent 5S1 conformation. rIDUA can be labeled with fluorescent Cy5 ABP 2, which allows monitoring of the delivery of therapeutic recombinant enzyme to lysosomes, as is intended in enzyme replacement therapy for the treatment of MPS I patients.

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Section: Discussionmentioning

confidence: 99%

New Irreversible α‐l‐Iduronidase Inhibitors and Activity‐Based Probes

Artola

Kuo

McMahon

et al. 2018

Chemistry A European J

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“…The first crystal structures of a member of the family of glycoside hydrolases have been recently determined (19,20). This protein, designated GH116 ␤-glucosidase from Thermoanaerobacterium xylanolyticum (Tx), shares overall 32% sequence identity with hGBA2 and ϳ40% in the catalytic domain, with excellent correspondence between the active-site residues.…”

Section: Orthologous Mutations In the Mouse Gba2 Gene Cause A Loss Ofmentioning

confidence: 99%

“…All residues that bind the sugar moiety in TxGH116 are conserved in hGBA2 (19). The homology to TxGH116 allows modeling of human GBA2, based on PDB accession code 5BVU (20) using the workspace modeling approach of the Swiss-Model suite (21). The model of hGBA2 encompasses residues 77-888, with ␤-D-glucose as a ligand superimposed from the crystal structure 5BX5 (19).…”

Section: Orthologous Mutations In the Mouse Gba2 Gene Cause A Loss Ofmentioning

confidence: 99%

Species-specific differences in nonlysosomal glucosylceramidase GBA2 function underlie locomotor dysfunction arising from loss-of-function mutations

Woeste

Stern

Raju

et al. 2019

Journal of Biological Chemistry

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The nonlysosomal glucosylceramidase ␤2 (GBA2) catalyzes the hydrolysis of glucosylceramide to glucose and ceramide. Mutations in the human GBA2 gene have been associated with hereditary spastic paraplegia (HSP), autosomal-recessive cerebellar ataxia (ARCA), and the Marinesco-Sjögren-like syndrome. However, the underlying molecular mechanisms are ill-defined. Here, using biochemistry, immunohistochemistry, structural modeling, and mouse genetics, we demonstrate that all but one of the spastic gait locus #46 (SPG46)-connected mutations cause a loss of GBA2 activity. We demonstrate that GBA2 proteins form oligomeric complexes and that proteinprotein interactions are perturbed by some of these mutations. To study the pathogenesis of GBA2-related HSP and ARCA in vivo, we investigated GBA2-KO mice as a mammalian model system. However, these mice exhibited a high phenotypic variance and did not fully resemble the human phenotype, suggesting that mouse and human GBA2 differ in function. Whereas some GBA2-KO mice displayed a strong locomotor defect, others displayed only mild alterations of the gait pattern and no signs of cerebellar defects. On a cellular level, inhibition of GBA2 activity in isolated cerebellar neurons dramatically affected F-actin dynamics and reduced neurite outgrowth, which has been associated with the development of neurological disorders. Our results shed light on the molecular mechanism underlying the pathogenesis of GBA2-related HSP and ARCA and reveal species-specific differences in GBA2 function in vivo.

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“…Meanwhile, cyclophellitol aziridine ABPs labeling -galactosidases, -glucosidases, -fucosidase, -galactosidases, and -glucuronidase have been successfully generated (160)(161)(162)(163)(164). These ABPs find several applications, such as quantitative detection and localization of glycosidases in cells and tissues, as well as identification and characterization of glycosidase inhibitors by competitive ABP profiling (165)(166)(167). Another important application lies in the demonstration of the deficiency of glycosidases in assisting the diagnosis of corresponding inherited diseases.…”

Section: Covalent Mechanism-based Inhibitors and Diagnostic Activity-mentioning

confidence: 99%

Distinguishing the differences in β-glycosylceramidase folds, dynamics, and actions informs therapeutic uses

Bdira

Artola

Overkleeft

et al. 2018

Journal of Lipid Research

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Glycoconjugates play essential roles in diverse biological processes and abnormalities and have been linked to multiple pathologies. The staggering structural diversity of glycoconjugates stems from the almost infinite possible combinations by which multiple monosaccharide building blocks may be linked with each other (1). For instance, 1,056 unique trisaccharides can be formed just from 3 different monosaccharides. Polysaccharides are very stable compounds: their spontaneous hydrolysis takes place at a rate of 10 15 s 1 , corresponding to a half-life of 4.7 million years (2). To allow for the efficient metabolism of glycoconjugates, enzymes have evolved as specialized catalysts. In most organisms, an estimated 1% to 3% of the genes encode carbohydrate-active enzymes (3). Among these are glycoside hydrolases (GHs) that can enhance the rate of hydrolysis of specific carbohydrate glycosidic bonds in glycoconjugates more than 10 17 times (2). These GH enzymes show marked specificity regarding number, position, and configuration of the hydroxyl groups in their substrate sugar. They are widely applied in biotechnology, for example, in biofuel production, paper pulp bleaching, and the food industry (4, 5). Likewise, specific GH inhibitors are extensively used as agrochemicals and therapeutic agents (6-8). Abnormalities in GHs underlie metabolic disorders in humans, for instance, inherited lysosomal storage disorders and lactose intolerance (9, 10). GH enzymes differ in substrate specificity, mode of enzymatic attack Abstract Glycosyl hydrolases (GHs) are carbohydrate-active enzymes that hydrolyze a specific -glycosidic bond in glycoconjugate substrates; -glucosidases degrade glucosylceramide, a ubiquitous glycosphingolipid. GHs are grouped into structurally similar families that themselves can be grouped into clans. GH1, GH5, and GH30 glycosidases belong to clan A hydrolases with a catalytic (/) 8 TIM barrel domain, whereas GH116 belongs to clan O with a catalytic (/) 6 domain. In humans, GH abnormalities underlie metabolic diseases. The lysosomal enzyme glucocerebrosidase (family GH30), deficient in Gaucher disease and implicated in Parkinson disease etiology, and the cytosol-facing membrane-bound glucosylceramidase (family GH116) remove the terminal glucose from the ceramide lipid moiety. Here, we compare enzyme differences in fold, action, dynamics, and catalytic domain stabilization by binding site occupancy. We also explore other glycosidases with reported glycosylceramidase activity, including human cytosolic -glucosidase, intestinal lactase-phlorizin hydrolase, and lysosomal galactosylceramidase. Last, we describe the successful translation of research to practice: recombinant glycosidases and glucosylceramide metabolism modulators are approved drug products (enzyme replacement therapies). Activity-based probes now facilitate the diagnosis of enzyme deficiency and screening for compounds that interact with the catalytic pocket of glycosidases. Future research may deepen the understanding of the function...

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1,6-Cyclophellitol Cyclosulfates: A New Class of Irreversible Glycosidase Inhibitor

Cited by 44 publications

References 40 publications

New Irreversible α‐l‐Iduronidase Inhibitors and Activity‐Based Probes

New Irreversible α‐l‐Iduronidase Inhibitors and Activity‐Based Probes

Species-specific differences in nonlysosomal glucosylceramidase GBA2 function underlie locomotor dysfunction arising from loss-of-function mutations

Distinguishing the differences in β-glycosylceramidase folds, dynamics, and actions informs therapeutic uses

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