title: Tool compounds robustly increase turnover of an artificial substrate by glucocerebrosidase in human brain lysates.
abstract: Mutations in glucocerebrosidase (GBA1) cause Gaucher disease and also represent a common risk factor for Parkinson's disease and Dementia with Lewy bodies. Recently, new tool molecules were described which can increase turnover of an artificial substrate 4MUG when incubated with mutant N370S GBA1 from human spleen. Here we show that these compounds exert a similar effect on the wild-type enzyme in a cell-free system. In addition, these tool compounds robustly increase turnover of 4MUG by GBA1 derived from human cortex, despite substantially lower glycosylation of GBA1 in human brain, suggesting that the degree of glycosylation is not important for compound binding. Surprisingly, these tool compounds failed to robustly alter GBA1 turnover of 4MUG in the mouse brain homogenate. Our data raise the possibility that in vivo models with humanized glucocerebrosidase may be needed for efficacy assessments of such small molecules.
authors.affiliation: Pfizer Neuroscience Research Unit, 610 Main Street, Cambridge, MA, United States of America.
authors.forename: Zdenek
authors.initials: Z
authors.lastname: Berger
authors.affiliation: Pfizer Neuroscience Research Unit, 610 Main Street, Cambridge, MA, United States of America.
authors.forename: Sarah
authors.initials: S
authors.lastname: Perkins
authors.affiliation: Pfizer Neuroscience Research Unit, 610 Main Street, Cambridge, MA, United States of America.
authors.forename: Claude
authors.initials: C
authors.lastname: Ambroise
authors.affiliation: Pfizer Neuroscience Research Unit, 610 Main Street, Cambridge, MA, United States of America.
authors.forename: Christine
authors.initials: C
authors.lastname: Oborski
authors.affiliation: Pfizer Structural Biology Group, Eastern Point Road, Groton, CT, United States of America.
authors.forename: Matthew
authors.initials: M
authors.lastname: Calabrese
authors.affiliation: Pfizer Primary Pharmacology Group, Eastern Point Road, Groton, CT, United States of America.
authors.forename: Stephen
authors.initials: S
authors.lastname: Noell
authors.affiliation: Pfizer Neuroscience Research Unit, 610 Main Street, Cambridge, MA, United States of America.
authors.forename: David
authors.initials: D
authors.lastname: Riddell
authors.affiliation: Pfizer Neuroscience Research Unit, 610 Main Street, Cambridge, MA, United States of America.
authors.forename: Warren D
authors.initials: WD
authors.lastname: Hirst
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology*; D002474:Cell-Free System; D005962:Glucosylceramidase / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D006923:Hymecromone / Q000031:analogs & derivatives* / Q000378:metabolism; D051379:Mice
chemical_list: D006923:Hymecromone; C043647:4-methylumbelliferyl-6-sulfo-2-acetamido-2-deoxy-beta-glucopyranoside; D005962:Glucosylceramidase