title: Potent and selective activity-based probes for GH27 human retaining α-galactosidases.
abstract: Lysosomal degradation of glycosphingolipids is mediated by the consecutive action of several glycosidases. Malfunctioning of one of these hydrolases can lead to a lysosomal storage disorder such as Fabry disease, which is caused by a deficiency in α-galactosidase A. Herein we describe the development of potent and selective activity-based probes that target retaining α-galactosidases. The fluorescently labeled aziridine-based probes 3 and 4 inhibit the two human retaining α-galactosidases αGal A and αGal B covalently and with high affinity. Moreover, they enable the visualization of the endogenous activity of both α-galactosidases in cell extracts, thereby providing a means to study the presence and location of active enzyme levels in different cell types, such as healthy cells versus those derived from Fabry patients.
authors.affiliation: Leiden Institute of Chemistry and The Netherlands Proteomics Centre, Leiden University , P.O. Box 9502, 2300 RA Leiden, The Netherlands.
authors.forename: Lianne I
authors.initials: LI
authors.lastname: Willems
authors.forename: Thomas J M
authors.initials: TJ
authors.lastname: Beenakker
authors.forename: Benjamin
authors.initials: B
authors.lastname: Murray
authors.forename: Saskia
authors.initials: S
authors.lastname: Scheij
authors.forename: Wouter W
authors.initials: WW
authors.lastname: Kallemeijn
authors.forename: Rolf G
authors.initials: RG
authors.lastname: Boot
authors.forename: Marri
authors.initials: M
authors.lastname: Verhoek
authors.forename: Wilma E
authors.initials: WE
authors.lastname: Donker-Koopman
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Ferraz
authors.forename: Erwin R
authors.initials: ER
authors.lastname: van Rijssel
authors.forename: Bogdan I
authors.initials: BI
authors.lastname: Florea
authors.forename: Jeroen D C
authors.initials: JD
authors.lastname: Codée
authors.forename: Gijsbert A
authors.initials: GA
authors.lastname: van der Marel
authors.forename: Johannes M F G
authors.initials: JM
authors.lastname: Aerts
authors.forename: Herman S
authors.initials: HS
authors.lastname: Overkleeft
mesh_terms: D001388:Aziridines / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D015394:Molecular Structure; D013329:Structure-Activity Relationship; D000519:alpha-Galactosidase / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D001388:Aziridines; D005456:Fluorescent Dyes; C033132:aziridine; D000519:alpha-Galactosidase