Activity-Based Probes for Glycosidases: Profiling and Other Applications

Kuo, C.J.; Meel, Eline van; Kytidou, Kassiani; Kallemeijn, Wouter W.; Witte, Martin D.; Overkleeft, Herman S.; Artola, Marta; Aerts, Johannes M. F. G.

doi:10.1016/bs.mie.2017.06.039

Cited by 21 publications

(22 citation statements)

References 44 publications

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“…Cyclophellitol aziridine ABPs labeling α-galactosidases, α-glucosidases, α-fucosidase, α-iduronidase, β-galactosidases, and β-glucuronidase, as well as cyclophellitol ABPs labelling galactocerebrosidase, were designed [60][61][62][63][64][65]. Applications of ABPs are the quantitative detection and localization of glycosidases in cells and tissues, as well as identification and characterization of glycosidase inhibitors by competitive ABP profiling [66,67].…”

Section: Gcase Protein and Life Cyclementioning

confidence: 99%

Glucocerebrosidase: Functions in and Beyond the Lysosome

Boer

Smeden

Bouwstra

et al. 2020

JCM

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Glucocerebrosidase (GCase) is a retaining β-glucosidase with acid pH optimum metabolizing the glycosphingolipid glucosylceramide (GlcCer) to ceramide and glucose. Inherited deficiency of GCase causes the lysosomal storage disorder named Gaucher disease (GD). In GCase-deficient GD patients the accumulation of GlcCer in lysosomes of tissue macrophages is prominent. Based on the above, the key function of GCase as lysosomal hydrolase is well recognized, however it has become apparent that GCase fulfills in the human body at least one other key function beyond lysosomes. Crucially, GCase generates ceramides from GlcCer molecules in the outer part of the skin, a process essential for optimal skin barrier property and survival. This review covers the functions of GCase in and beyond lysosomes and also pays attention to the increasing insight in hitherto unexpected catalytic versatility of the enzyme.

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Section: Gcase Protein and Life Cyclementioning

confidence: 99%

Glucocerebrosidase: Functions in and Beyond the Lysosome

Boer

Smeden

Bouwstra

et al. 2020

JCM

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“…Thus, the difference in phenotype cannot be explained by compensation through an increase in GBA1 activity. Another way of determining GBA2 activity would be to use activity-based probes, which can be used on live cells and brain sections (9,47). Whether also other risk factors contribute to disease progression in SPG46 patients remains unknown.…”

Section: Gba2 Mutations and Locomotor Dysfunctionmentioning

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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“…Here, to address these questions, we quantitatively characterized the delivery of axonal degradative lysosomes by applying four enzymatic-activity-based fluorescent probes that selectively bind to active lysosomal hydrolases, including cathepsins D, B, and L and glucocerebrosidase (GCase) (Chen et al, 2000; Creasy et al, 2007; Kuo et al, 2018). By time-lapse imaging of cortical neurons cultured in microfluidic devices that allow physical and fluidic separation of axons from cell bodies and dendrites, we reveal that enzymatically active degradative lysosomes labeled within the soma chamber can influx into distal axons and target to autophagosomes and α-synuclein cargos for local degradation.…”

Section: Introductionmentioning

confidence: 99%

Neuronal Soma-Derived Degradative Lysosomes Are Continuously Delivered to Distal Axons to Maintain Local Degradation Capacity

et al. 2019

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SUMMARY Neurons face the challenge of maintaining cellular homeostasis through lysosomal degradation. While enzymatically active degradative lysosomes are enriched in the soma, their axonal trafficking and positioning and impact on axonal physiology remain elusive. Here, we characterized axon-targeted delivery of degradative lysosomes by applying fluorescent probes that selectively label active forms of lysosomal cathepsins D, B, L, and GCase. By time-lapse imaging of cortical neurons in microfluidic devices and standard dishes, we reveal that soma-derived degradative lysosomes rapidly influx into distal axons and target to autophagosomes and Parkinson disease-related α-synuclein cargos for local degradation. Impairing lysosome axonal delivery induces an aberrant accumulation of autophagosomes and α-synuclein cargos in distal axons. Our study demonstrates that the axon is an active compartment for local degradation and reveals fundamental aspects of axonal lysosomal delivery and maintenance. Our work establishes a foundation for investigations into axonal lysosome trafficking and functionality in neurodegenerative diseases.

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Activity-Based Probes for Glycosidases: Profiling and Other Applications

Cited by 21 publications

References 44 publications

Glucocerebrosidase: Functions in and Beyond the Lysosome

Glucocerebrosidase: Functions in and Beyond the Lysosome

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

Neuronal Soma-Derived Degradative Lysosomes Are Continuously Delivered to Distal Axons to Maintain Local Degradation Capacity

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