8r1m
Structure of TxGH116 with covalently bound N-azido-octyl aziridine
Structural highlights
FunctionPublication Abstract from PubMedGBA2, the non-lysosomal beta-glucosylceramidase, is an enzyme involved in glucosylceramide metabolism. Pharmacological inhibition of GBA2 by N-alkyl iminosugars is well tolerated and benefits patients suffering from Sandhoff and Niemann-Pick type C diseases, and GBA2 inhibitors have been proposed as candidate-clinical drugs for the treatment of parkinsonism. With the ultimate goal to unravel the role of GBA2 in (patho)physiology, we sought to develop a GBA2-specific activity-based probe (ABP). A library of probes was tested for activity against GBA2 and the two other cellular retaining beta-glucosidases, lysosomal GBA1 and cytosolic GBA3. We show that beta-d-arabinofuranosyl cyclitol aziridine (beta-d-Araf aziridine) reacts with the GBA2 active site nucleophile to form a covalent and irreversible bond. Fluorescent beta-d-Araf aziridine probes potently and selectively label GBA2 both in vitro and in cellulo, allowing for visualization of the localization of overexpressed GBA2 using fluorescence microscopy. Co-staining with an antibody selective for the lysosomal beta-glucosylceramidase GBA1, shows distinct subcellular localization of the two enzymes. We proffer our ABP technology for further delineating the role and functioning of GBA2 in disease and propose the beta-d-Araf aziridine scaffold as a good starting point for the development of GBA2-specific inhibitors for clinical development. Selective labelling of GBA2 in cells with fluorescent beta-d-arabinofuranosyl cyclitol aziridines.,Su Q, Louwerse M, Lammers RF, Maurits E, Janssen M, Boot RG, Borlandelli V, Offen WA, Linzel D, Schroder SP, Davies GJ, Overkleeft HS, Artola M, Aerts JMFG Chem Sci. 2024 Sep 3;15(37):15212-20. doi: 10.1039/d3sc06146a. PMID:39246358[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
| ||||||||||||||||||||
