Change of isoforms' spectra of α-l-fucosidase from human skin fibroblasts in intracellular storage of nonhydrolyzable substances

Abstract

The effect of exogenous and endogenous products storage in lysosomes on the activity and multiple forms of alpha-L-fucosidase from human skin fibroblasts was investigated. It was shown that sucrose load, modelling intralysosomal accumulation of nonhydrolyzable products, causes certain changes in secretion level of alpha-L-fucosidase and multiple forms' spectra of the intracellular and secreted enzymes. These changes were different for the enzyme from embryonal and postnatal normal fibroblasts. Some changes of secreted alpha-L-fucosidase isoforms' spectra were found in fibroblasts from a patient with Fabry's disease, characterized by the intralysosomal storage of di- and trihexosylceramides. The alterations of isoforms' profiles in Fabry fibroblasts at the early and late accumulation stages were similar to those in sucrose-loaded embryonal and postnatal fibroblasts, respectively. It is proposed that intralysosomal accumulation of nonhydrolyzable compounds influences the alpha-L-fucosidase posttranslational processing.

Changes on the Caco-2 Secretome through Differentiation Analyzed by 2-D Differential In-Gel Electrophoresis (DIGE)

“…To study the extracellular medium, cells were kept in FCS-free medium for 24 h before collection [62]. After 24 h, medium was collected and centrifuged at 600 g for 5 min, so that particles on suspension were discarded with the pellet.…”