Novel Activity‐Based Probes for Broad‐Spectrum Profiling of Retaining β‐Exoglucosidases In Situ and In Vivo

Kallemeijn, Wouter W.; Li, Kah‐Yee; Witte, Martin D.; Marques, André R. A.; Aten, Jan; Scheij, Saskia; Jiang, Jianbing; Willems, Lianne I.; Voorn-Brouwer, Tineke; Roomen, Cindy P. A. A. van; Ottenhoff, Roelof; Boot, Rolf G.; Elst, Hans van den; Walvoort, Marthe T. C.; Florea, Bogdan I.; Codée, Jeroen D. C.; Marel, Gijsbert A. van der; Aerts, Johannes M. F. G.; Overkleeft, Herman S.

doi:10.1002/anie.201207771

Cited by 109 publications

(98 citation statements)

References 28 publications

(3 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…ABPP probes for glucosidases, such as glucocerebrosidase, comprise epoxide and aziridine-containing cyclophellitol mimics (Figure 4i) [66,67]. The SE electrophile (Figure 4j) can also modify aspartates and glutamates that act as active-site proton donors/acceptors in dienoyl-CoA isomerase and long-chain specific acylCoA dehydrogenase [6 ].…”

Section: Covalent Modification Of Aspartate and Glutamatementioning

confidence: 99%

Covalent protein modification: the current landscape of residue-specific electrophiles

Shannon

Weerapana

2015

Current Opinion in Chemical Biology

198

178

View full text Add to dashboard Cite

Section: Covalent Modification Of Aspartate and Glutamatementioning

confidence: 99%

Covalent protein modification: the current landscape of residue-specific electrophiles

Shannon

Weerapana

2015

Current Opinion in Chemical Biology

198

178

View full text Add to dashboard Cite

“…(a) Structural formulas of competitive, reversible inhibitor isofagomine (IFG 1 ), irreversible inhibitors conduritol β-epoxide (CBE 2 ), semi-irreversible inhibitor 2-deoxy-2-fluoro-β- d -glucopyranosyl- N -phenyltrifluoroacetimidate 33 (fluoro 3 ), cyclophellitol β-epoxide type ABP 4 (MDW933, green fluorescent), and β-epoxide-type ABP 5 (MDW941, red fluorescent). 35 (b) Irreversible binding mechanism of β-epoxide-type ABPs to the nucleophile of GBA via its double-displacement mechanism. (c) Hydrolysis of fluoro 3 and temporary trapping of the glycosylated nucleophile adducts of GBA.…”

mentioning

confidence: 99%

Stabilization of Glucocerebrosidase by Active Site Occupancy

Bdira¹,

Kallemeijn²,

Oussoren³

et al. 2017

ACS Chem. Biol.

Self Cite

View full text Add to dashboard Cite

Glucocerebrosidase (GBA) is a lysosomal β-glucosidase that degrades glucosylceramide. Its deficiency results in Gaucher disease (GD). We examined the effects of active site occupancy of GBA on its structural stability. For this, we made use of cyclophellitol-derived activity-based probes (ABPs) that bind irreversibly to the catalytic nucleophile (E340), and for comparison, we used the potent reversible inhibitor isofagomine. We demonstrate that cyclophellitol ABPs improve the stability of GBA in vitro, as revealed by thermodynamic measurements (Tm increase by 21 °C), and introduce resistance to tryptic digestion. The stabilizing effect of cell-permeable cyclophellitol ABPs is also observed in intact cultured cells containing wild-type GBA, N370S GBA (labile in lysosomes), and L444P GBA (exhibits impaired ER folding): all show marked increases in lysosomal forms of GBA molecules upon exposure to ABPs. The same stabilization effect is observed for endogenous GBA in the liver of wild-type mice injected with cyclophellitol ABPs. Stabilization effects similar to those observed with ABPs were also noted at high concentrations of the reversible inhibitor isofagomine. In conclusion, we provide evidence that the increase in cellular levels of GBA by ABPs and by the reversible inhibitor is in part caused by their ability to stabilize GBA folding, which increases the resistance of GBA against breakdown by lysosomal proteases. These effects are more pronounced in the case of the amphiphilic ABPs, presumably due to their high lipophilic potential, which may promote further structural compactness of GBA through hydrophobic interactions. Our study provides further rationale for the design of chaperones for GBA to ameliorate Gaucher disease.

show abstract

“…For example, xyloglucan endotransglycosylase activity has been monitored in situ by infiltrating natural fluorogenic substrates for that enzyme [61]. Furthermore, cell-permeable fluorescent activity-based probes for glycosidases can be used to locate active enzymes in situ [62].…”

Section: Exciting Directions For Future Bgal Researchmentioning

confidence: 99%

Beta galactosidases in Arabidopsis and tomato–a mini review

Chandrasekar

Hoorn

2016

Biochemical Society Transactions

View full text Add to dashboard Cite

Beta galactosidases (BGALs) are glycosyl hydrolases that remove terminal β-D-galactosyl residues from β-D-galactosides. There are 17 predicted BGAL genes in the genomes of both Arabidopsis (BGAL1-17) and tomato (TBG1-17). All tested BGALs have BGAL activity but their distinct expression profiles and ancient phylogenetic separation indicates that these enzymes fulfil diverse, non-redundant roles in plant biology. The majority of these BGALs are predicted to have signal peptide and thought to act during cell wall-related biological processes. Interestingly, deletion of BGAL6 and BGAL10 in Arabidopsis causes reduced mucilage release during seed imbibition and shorter siliques respectively, whereas TBG4 depletion by RNAi decreases in fruit softening in tomato. The majority of plant BGALs remain to be characterized.

show abstract

Novel Activity‐Based Probes for Broad‐Spectrum Profiling of Retaining β‐Exoglucosidases In Situ and In Vivo

Cited by 109 publications

References 28 publications

Covalent protein modification: the current landscape of residue-specific electrophiles

Covalent protein modification: the current landscape of residue-specific electrophiles

Stabilization of Glucocerebrosidase by Active Site Occupancy

Beta galactosidases in Arabidopsis and tomato–a mini review

Contact Info

Product

Resources

About