Pathobiochemistry of the Fabry disease and other sphingolipidoses with α-galaktosidase A dysfunction

Fabry disease is an inherited defect of lysosomal α-galactosidase A (α-GALA), causing progressive accumulation of glycosphingolipids with terminal α-galactosyl moieties, especially globotriaosylceramide (Gb3Cer) and in to a small extent also galabiosylceramide (Ga2Cer) and blood group B glycolipids, in most tissues and body fluids. This diploma thesis is an extension of previous laboratory studies and intends to contribute to clarification of some specific features of catabolic pathways of glycolipids substrates in lysosomal storage disorders, especially blood group B glycolipids. Therefore, analysis of human pancreas and lungs tissues was performed using TLC imunodetection and immunohistochemical analysis of these glycolipids. The most striking observation was massive accumulation of B-6-2 glycolipid and of others complex B-glycolipids in the pancreas of the patient with Fabry disease with blood group B. The level of blood group B substrates exceeded significantly storage of Gb3Cer substrate. An important part of this work were metabolic experiments in cell cultures in order to answer the question about participation of related glycosidases - α-galactosidase A and α-N- acetylgalactosaminidase (α -NAGA) in the lysosomal degradation of glycosphingolipids with terminal α-galactose. Loading experiments were..