title: Cell surface associated glycohydrolases in normal and Gaucher disease fibroblasts.
abstract: Gaucher disease (GD) is the most common lysosomal disorder and is caused by an inherited autosomal recessive deficiency in β-glucocerebrosidase. This enzyme, like other glycohydrolases involved in glycosphingolipid (GSL) metabolism, is present in both plasma membrane (PM) and intracellular fractions. We analyzed the activities of CBE-sensitive β-glucosidase (GBA1) and AMP-DNM-sensitive β-glucosidase (GBA2) in total cell lysates and PM of human fibroblast cell lines from control (normal) subjects and from patients with GD clinical types 1, 2, and 3. GBA1 activities in both total lysate and PM of GD fibroblasts were low, and their relative percentages were similar to those of control cells. In contrast, GBA2 activities were higher in GD cells than in control cells, and the degree of increase differed among the three GD types. The increase of GBA2 enzyme activity was correlated with increased expression of GBA2 protein as evaluated by QRT-PCR. Activities of β-galactosidase and β-hexosaminidase in PM were significantly higher for GD cells than for control cells and also showed significant differences among the three GD types, suggesting the occurrence of cross-talk among the enzymes involved in GSL metabolism. Our findings indicate that the profiles of glycohydrolase activities in PM may provide a valuable tool to refine the classification of GD into distinct clinical types.
authors.affiliation: Department of Medical Chemistry, Biochemistry and Biotechnology, University of Milan, 20090, Segrate, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Aureli
authors.forename: Rosaria
authors.initials: R
authors.lastname: Bassi
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Loberto
authors.forename: Stefano
authors.initials: S
authors.lastname: Regis
authors.forename: Alessandro
authors.initials: A
authors.lastname: Prinetti
authors.forename: Vanna
authors.initials: V
authors.lastname: Chigorno
authors.forename: Johannes M
authors.initials: JM
authors.lastname: Aerts
authors.forename: Rolf G
authors.initials: RG
authors.lastname: Boot
authors.forename: Mirella
authors.initials: M
authors.lastname: Filocamo
authors.forename: Sandro
authors.initials: S
authors.lastname: Sonnino
mesh_terms: D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D005347:Fibroblasts / Q000201:enzymology; D005776:Gaucher Disease / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D005962:Glucosylceramidase / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006801:Humans; D001617:beta-Glucosidase / Q000378:metabolism
chemical_list: D006026:Glycoside Hydrolases; D001617:beta-Glucosidase; C517537:GBA2 protein, human; D005962:Glucosylceramidase