Study of the Hurler syndrome using cell culture: definition of the biochemical phenotype and the effect of ascorbic acid on the mutant cell

Schafer, Irwin A.; Sullivan, Julia C.; Kofoed, J. A.; Robertson, W. W.

doi:10.1172/jci105727

Cited by 31 publications

(8 citation statements)

References 25 publications

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“…The observed affinity for GAGs is thus biologically relevant because the CPP concentration generally required for efficient cell penetration (1-10 mM) and physiological GAG concentration are of similar magnitude. For fibroblasts, for example, 2 mg of sulfated GAGs/mg dry tissue are reported (27), corresponding to a macroscopic GAG concentration of ;7 mM using a typical molecular weight of GAGs found in fibroblasts (28) and an average tissue water content. The effective concentration of individual binding sites is even higher because each GAG can bind several CPPs (Table 1).…”

Section: Binding Affinitymentioning

confidence: 99%

Binding and Clustering of Glycosaminoglycans: A Common Property of Mono- and Multivalent Cell-Penetrating Compounds

Ziegler

Seelig

2008

Biophysical Journal

116

109

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Recent observations in cell culture provide evidence that negatively charged glycosaminoglycans (GAGs) at the surface of biological cells bind cationic cell-penetrating compounds (CPCs) and cluster during CPC binding, thereby contributing to their endocytotic uptake. The GAG binding and clustering occur in the low-micromolar concentration range and suggest a tight interaction between GAGs and CPCs, although the relation between binding affinity and specificity of this interaction remains to be investigated. We therefore measured the GAG binding and clustering of various mono- and multivalent CPCs such as DNA transfection vectors (polyethylenimine; 1,2-dioleoyl-3-trimethylammonium-propane), amino acid homopolymers (oligoarginine; oligolysine), and cell-penetrating peptides (Penetratin; HIV-1 Tat) by means of isothermal titration calorimetry and dynamic light scattering. We find that these structurally diverse CPCs share the property of GAG binding and clustering. The binding is very tight (microscopic dissociation constants between 0.34 and 1.34 microM) and thus biologically relevant. The hydrodynamic radius of the resulting aggregates ranges from 78 nm to 586 nm, suggesting that they consist of numerous GAG chains cross-linked by CPCs. Likewise, the membrane-permeant monovalent cation acridine orange leads to GAG binding and clustering, in contrast to its membrane-impermeant structural analogs propidium iodide and ethidium bromide. Because the binding and clustering of GAGs were found to be a common denominator of all CPCs tested, these properties might be helpful to identify further CPCs.

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Section: Binding Affinitymentioning

confidence: 99%

Binding and Clustering of Glycosaminoglycans: A Common Property of Mono- and Multivalent Cell-Penetrating Compounds

Ziegler

Seelig

2008

Biophysical Journal

116

109

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“…In the course of studies on Vol. 151 dermatan sulphate-storage diseases (Hurler's and Hunter's syndromes) partial chemical characterization of intracellular as well as extracellular dermatan sulphate from normal cells has been performed (Matalon & Dorfman, 1966;Schafer et al, 1968;DiFerrante et al, 1971). Moreover, the kinetics of intracellular accumulation and secretion into the medium has been documented (Fratantoni et al, 1968;DiFerrante et al, 1971).…”

mentioning

confidence: 99%

The copolymeric structure of dermatan sulphate produced by cultured human fibroblasts. Different distribution of iduronic acid- and glucuronic acid-containing units in soluble and cell-associated glycans

Malström

Carlstedt

Åberg

et al. 1975

Biochemical Journal

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The structure of dermatan [35S]sulphate-chondroitin [35S]sulphate copolymers synthesized and secreted by fibroblasts in culture was studied. 35S-labelled glycosaminoglycans were isolated from the medium, a trypsin digest of the cells and the cell residue after 72h of 35SO42-incorporation. The galactosaminoglycan component (dermatan sulphatechondroitin sulphate copolymers) was isolated and subjected to various degradation procedures including digestion with testicular hyaluronidase, chondroitinase-AC and-ABC and periodate oxidation followed by alkaline elimination. The galactosaminoglycans from the various sources displayed significant structural differences with regard to the distribution of various repeating units, i.e. IdUA-GalNAc-SO4 (L-iduronic acid-N-acetyl-galactosamine sulphate), GlcUA-GalNAc-SO4 (D-glucuronic acid-N-acetylgalactosamine-sulphate) and IdUA(-SO4)-GalNAc (L-iduronosulphate-N-acetylgalactosamine). The galactosaminoglycans of the cell residue contained larger amounts of IdUA-GalNAc-SO4 than did those isolated from the medium or those released by trypsin. In contrast, the glycans from the latter 2 sources contained large proportions of periodate-resistant repeat periods [GlcUA-GalNAc-SO4 and IdUA(-SO4)-GalNAc]. Periods containing L-iduronic acid sulphate were particularly prominent in copolymers found in the medium. Kinetic studies indicated that the 35S-labelled glycosaminoglycan of the cell residue accumulated radioactivity more slowly than did the glycans of other fractions, indicating that the material remaining with the cells was not exclusively a precursor of the secreted polymers. The presence of copolymers rich in glucuronic acid or iduronic acid sulphate residues in the soluble fractions may be the result of selective secretion from the cells. Alternatively, extracellular, polymer-level modifications such as C-5 inversion of L-iduronic acid to D-glucuronic acid, or sulphate rearrangements, would yield similar results.

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“…Nevertheless, a fibroblast ctilUne can be carried otit from a skin fragment, and in this way tbe disease can be deeisively typed by cytocbemical, bioehetnical, enzymatic and genetic means (Danes & Beam 1966, Matalon & Dorfman 1966, Schafer et al 1968, Tondeur et al 1970. Nevertheless, a fibroblast ctilUne can be carried otit from a skin fragment, and in this way tbe disease can be deeisively typed by cytocbemical, bioehetnical, enzymatic and genetic means (Danes & Beam 1966, Matalon & Dorfman 1966, Schafer et al 1968, Tondeur et al 1970.…”

Section: Dfscusslonmentioning

confidence: 99%

“…The type of MPS or lipidose cannot be determined by electron mieioscopy alone. Nevertheless, a fibroblast ctilUne can be carried otit from a skin fragment, and in this way tbe disease can be deeisively typed by cytocbemical, bioehetnical, enzymatic and genetic means (Danes & Beam 1966, Matalon & Dorfman 1966, Schafer et al 1968, Tondeur et al 1970. Tbus, even clinically normal skin mtisl be biopsied if the diagnosis of MPS is to be established.…”

Section: Dfscusslonmentioning

confidence: 99%

The Diagnosis of Mucopolysaccharidoses by Electron Microscopy of Skin Biopsies

et al. 1975

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An ultrastructural examination was carried out on the skin of six children suffering from Mucopolysaccharidosis I (MPSI or Hurler's disease) and MPS II (Hunter's disease). Both intracellular mucopolysaccharides and lipids were observed. The dermal cells, i.e. fibroblasts, macrophages, were loaded with multiple vacuolar inclusions thought to be of lysosomal origin. They appeared either content-free or filled with granular, fuzzy and/or pseudomyelinic structures. Identical abnormalities were observed within the Schwann cells, smooth muscle cells and keratinocytes. Mast cells showed peculiar "worm-like" inclusions apart from their normal granulations. Since ultrastructure of a skin sample may provide as much data as brain, liver or kidney, cutaneous electron microscopy can be recommended to confirm a diagnosis of MPS.

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Study of the Hurler syndrome using cell culture: definition of the biochemical phenotype and the effect of ascorbic acid on the mutant cell

Cited by 31 publications

References 25 publications

Binding and Clustering of Glycosaminoglycans: A Common Property of Mono- and Multivalent Cell-Penetrating Compounds

Binding and Clustering of Glycosaminoglycans: A Common Property of Mono- and Multivalent Cell-Penetrating Compounds

The copolymeric structure of dermatan sulphate produced by cultured human fibroblasts. Different distribution of iduronic acid- and glucuronic acid-containing units in soluble and cell-associated glycans

The Diagnosis of Mucopolysaccharidoses by Electron Microscopy of Skin Biopsies

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