title: Biochemical characterization of arylsulfatase E and functional analysis of mutations found in patients with X-linked chondrodysplasia punctata.
abstract: X-linked chondrodysplasia punctata (CDPX) is a congenital disorder characterized by abnormalities in cartilage and bone development. Mutations leading to amino acid substitutions were identified recently in CDPX patients, in the coding region of the arylsulfatase E (ARSE) gene, a novel member of the sulfatase gene family. Transfection of the ARSE full-length cDNA, in Cos7 cells, allowed us to establish that its protein product is a 60-kD precursor, which is subject to N-glycosylation, to give a mature 68-kD form that, unique among sulfatases, is localized to the Golgi apparatus. Five missense mutations found in CDPX patients were introduced into wild-type ARSE cDNA by site-directed mutagenesis. These mutants were transfected into Cos7 cells, and the arylsulfatase activity and biochemical properties were determined, to study the effect of these substitutions on the ARSE protein. One of the mutants behaves as the wild-type protein. All four of the other mutations resulted in a complete lack of arylsulfatase activity, although the substitutions do not appear to affect the stability and subcellular localization of the protein. The loss of activity due to these mutations confirms their involvement in the clinical phenotype and points to the importance of these residues in the correct folding of a catalytically active ARSE enzyme.
authors.affiliation: Telethon Institute of Genetics and Medicine, San Raffaele Biomedical Science Park, Milan, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Daniele
authors.forename: G
authors.initials: G
authors.lastname: Parenti
authors.forename: M
authors.initials: M
authors.lastname: d'Addio
authors.forename: G
authors.initials: G
authors.lastname: Andria
authors.forename: A
authors.initials: A
authors.lastname: Ballabio
authors.forename: G
authors.initials: G
authors.lastname: Meroni
mesh_terms: D000818:Animals; D001192:Arylsulfatases / Q000235:genetics*; D019556:COS Cells; D002806:Chondrodysplasia Punctata / Q000201:enzymology / Q000235:genetics*; D008040:Genetic Linkage; D006801:Humans; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D013347:Subcellular Fractions / Q000201:enzymology; D014162:Transfection; D014960:X Chromosome
chemical_list: C110155:ARSL protein, human; D001192:Arylsulfatases