Multiple sulfatase deficiency: catalytically inactive sulfatases are expressed from retrovirally introduced sulfatase cDNAs.

Abstract

Multiple sulfatase deficiency (MSD) is an inherited lysosomal storage disease characterized by the deficiency of at least seven sulfatases. The basic defect in MSD is thought to be in a post-translational modification common to all sulfatases. In accordance with this concept, RNAs of normal size and amount were detected in MSD fibroblasts for three sulfatases tested. cDNAs encoding arylsulfatase A, arylsulfatase B, or steroid sulfatase were introduced into MSD fibroblasts and fibroblasts with a single sulfatase deficiency by retroviral gene transfer. Infected fibroblasts overexpressed the respective sulfatase polypeptides. While in single-sulfatasedeficiency fibroblasts a concomitant increase of sulfatase activities was observed, MSD fibroblasts expred sulfatase polypeptides with a severely diminished catalytic activity.From these results we conclude that the mutation in MSD severely decreases the capacity of a co-or post-translational process that renders sulfatases enzymatically active or prevents their premature inactivation.

Sphingolipids—Their Metabolic Pathways and the Pathobiochemistry of Neurodegenerative Diseases

“…The primary defect of multiple sulfatase deficiency is not known, however, since expression of sulfatase cDNAs in fibroblasts of MSD patients leads to enzyme proteins with reduced catalytic activity, von Figura and co-workers have concluded that a defective co-or posttranslational modification is the cause of MSD. [339] They were able to detect a defective protein modification in arylsulfatases A and B in MSD cells. This was the transformation of a cysteine into a formylglycine residue.…”

Expression, Localization, Structural, and Functional Characterization of pFGE, the Paralog of the Cα-Formylglycine-generating Enzyme

“…Activity Assays for Sulfatases and FGE-Activity assays of FGE, steroid sulfatase, and galactose 6-sulfatase were performed as described earlier (14,22,23). For Western blot analysis, steroid sulfatase antiserum, galactose 6-sulfatase monoclonal antibodies (provided by Transkaryotic Therapies Inc., Cambridge, MA), and HA tag monoclonal antibodies (see above) were used as primary antibodies, which were detected with secondary horseradish peroxidase-conjugated goat antirabbit or anti-mouse antibodies.…”

Rapid degradation of an active formylglycine generating enzyme variant leads to a late infantile severe form of multiple sulfatase deficiency

“…10,11 Expression studies Cell culturing of patient skin fibroblasts and HT-1080 fibrosarcoma cells as well as stable transfection with pSB-FGE-His plasmids were described earlier. 6,7 The FGE-E130D mutation was created by site-directed mutagenesis according to the QuikChange Protocol (Stratagene, La Jolla, CA, USA) with Pfu polymerase and complementary primer pairs (coding sequence only: FGE_ E130D-His_c 5 0 -CATGGATGCCTATGATGTCAGTAATACTG-3 0 ).…”