title: Defective glucuronic acid transport from lysosomes of infantile free sialic acid storage disease fibroblasts.
abstract: Separation by h.p.l.c. and pulsed amperometric detection were employed to measure glucuronic acid (GlcUA) and other acidic monosaccharides in fibroblasts from patients with infantile free sialic acid storage disease (ISSD) and Salla disease. These lysosomal storage disorders result from defective carrier-mediated transport of free N-acetylneuraminic acid (NeuAc) out of cellular lysosomes. Three Salla disease fibroblast strains stored approx. 0.4 nmol of free GlcUA/mg of cell protein, whereas four ISSD strains stored approx. 5 nmol GlcUA/mg (normal is undetectable). The GlcUA content of the mutant cell strains, which by differential centrifugation and Percoll gradient fractionation was localized to the lysosomes, averaged 5% of the free NeuAc content of the cells. N-Glycolylneuraminic acid (NeuGc) also accumulated in ISSD cells, but only when they were grown in the presence of fetal calf serum, which contains abundant NeuGc. No other acidic monosaccharides were detected in any of the mutant cell strains. GlcUA egress studies revealed that 56% of the initial GlcUA content was lost from normal granular fractions after 2 min at 37 degrees C. For similarly loaded ISSD granular fractions, virtually no GlcUA was lost even after 6 min. The results indicate that GlcUA is recognized and transported by the lysosomal NeuAc carrier, and that GlcUA transport is impaired in the lysosomal disorders of free NeuAc storage.
authors.affiliation: Section of Human Biochemical Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.
authors.forename: H J
authors.initials: HJ
authors.lastname: Blom
authors.forename: H C
authors.initials: HC
authors.lastname: Andersson
authors.forename: R
authors.initials: R
authors.lastname: Seppala
authors.forename: F
authors.initials: F
authors.lastname: Tietze
authors.forename: W A
authors.initials: WA
authors.lastname: Gahl
mesh_terms: D001692:Biological Transport; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism; D005965:Glucuronates / Q000032:analysis / Q000493:pharmacokinetics*; D020723:Glucuronic Acid; D006801:Humans; D008247:Lysosomes / Q000378:metabolism*; D008661:Metabolism, Inborn Errors / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D009438:Neuraminic Acids / Q000032:analysis / Q000493:pharmacokinetics; D012794:Sialic Acids / Q000378:metabolism*; D013400:Sugar Acids / Q000032:analysis
chemical_list: D005965:Glucuronates; D009438:Neuraminic Acids; D012794:Sialic Acids; D013400:Sugar Acids; C032592:N-glycolylneuraminic acid; D020723:Glucuronic Acid