Impairment of the Golgi GDP-l-Fucose Transport and Unresponsiveness to Fucose Replacement Therapy in LAD II Patients

Sturla, Laura; Puglielli, Luigi; Tonetti, Michela; Berninsone, Patricia M.; Hirschberg, Carlos B.; Flora, Antonio De; Etzioni, Amos

doi:10.1203/00006450-200104000-00016

Cited by 63 publications

(54 citation statements)

References 35 publications

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“…No toxicity, determined by MTT (3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, was observed in these incubation conditions. Incorporation of the radiolabel in macromolecules was determined after PTA (phosphotungstic acid)/HCl precipitation, as previously described (36). Protein concentration was analyzed by DC Protein Assay (Bio-Rad).…”

Section: Methodsmentioning

confidence: 99%

“…However, the fact that some fucosylated antigens are undetectable (i.e. H antigen, selectin ligands), while LAD II cells incorporate low, but detectable levels of fucose into macromolecules, suggested that not all fucose-containing glycans are comparably decreased (36). The aim of the present study was the characterization of the residual fucose-containing glycoconjugates in cultured fibroblasts from a LAD II patient of Arab origin, in order to determine how the decreased availability of GDP-L-fucose inside the Golgi can affect their type and composition.…”

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confidence: 99%

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Differential Terminal Fucosylation of N-Linked Glycans Versus Protein O-Fucosylation in Leukocyte Adhesion Deficiency Type II (CDG IIc)

Sturla

Rampal

Haltiwanger

et al. 2003

Journal of Biological Chemistry

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LAD II/CDG IIc is a rare autosomal recessive disease characterized by a decreased expression of fucosylated antigens on cell surfaces that results in leukocyte adhesion deficiency and severe neurological and developmental abnormalities. Its molecular basis has been identified as a defect in the transporter of GDP-L-fucose into the Golgi lumen, which reduces the availability of the substrate for fucosyltransferases. During metabolic radiolabeling experiments using [ 3 H]fucose, LAD II fibroblasts incorporated significantly less radiolabel compared with control cells. However, fractionation and analysis of the different classes of glycans indicated that the decrease in [ 3 H]fucose incorporation is not generalized and is mainly confined to terminal fucosylation of N-linked oligosaccharides. In contrast, the total levels of protein O-fucosylation, including that observed in Notch protein, were unaffected. This finding demonstrates that the decrease in GDP-L-fucose levels in the fibroblast Golgi caused by the LAD II defect does not impair bulk protein O-fucosylation, but severely affects the bulk addition of fucose as a terminal modification of N-linked glycans. These data suggest that the severe clinical abnormalities including neurological and developmental ones observed in at least some of the LAD II patients may be related to alteration in recognition systems involving terminal fucose modifications of N-glycans and not be due to a defective O-fucosylation of proteins such as Notch.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

Differential Terminal Fucosylation of N-Linked Glycans Versus Protein O-Fucosylation in Leukocyte Adhesion Deficiency Type II (CDG IIc)

Sturla

Rampal

Haltiwanger

et al. 2003

Journal of Biological Chemistry

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“…In LADII, the GDP-D-mannose 4,6 dehydratase, an enzyme involved in de novo GDP-fucose synthesis from GDP-mannose, is normal in sequence and expression level (6,7). GDPfucose synthesis from free fucose via a so-called salvage pathway also appears to be normal in LADII patients (8). Thus, the most attractive model for a molecular defect in fucosylation involves decreased availability of GDP-fucose within the Golgi lumen.…”

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confidence: 99%

Golgi nucleotide sugar transport and leukocyte adhesion deficiency II

Hirschberg

2001

J. Clin. Invest.

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“…Interestingly, several patients of ArabIsraeli origin carry a different point mutation (T308R) (52). Analysis of the biochemical properties of the mutated GDP-fuc transporter in these patients revealed that it had a lower V max , but equal K m compared to the wild-type transporter (53).…”

Section: Nucleotide-sugar Transporters and Human Diseasementioning

confidence: 99%

Nucleotide-sugar transporters: structure, function and roles in vivo

Handford¹,

Rodriguez-Furlán²,

Orellana³

2006

Braz J Med Biol Res

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The glycosylation of glycoconjugates and the biosynthesis of polysaccharides depend on nucleotide-sugars which are the substrates for glycosyltransferases. A large proportion of these enzymes are located within the lumen of the Golgi apparatus as well as the endoplasmic reticulum, while many of the nucleotide-sugars are synthesized in the cytosol. Thus, nucleotide-sugars are translocated from the cytosol to the lumen of the Golgi apparatus and endoplasmic reticulum by multiple spanning domain proteins known as nucleotide-sugar transporters (NSTs). These proteins were first identified biochemically and some of them were cloned by complementation of mutants. Genome and expressed sequence tag sequencing allowed the identification of a number of sequences that may encode for NSTs in different organisms. The functional characterization of some of these genes has shown that some of them can be highly specific in their substrate specificity while others can utilize up to three different nucleotidesugars containing the same nucleotide. Mutations in genes encoding for NSTs can lead to changes in development in Drosophila melanogaster or Caenorhabditis elegans, as well as alterations in the infectivity of Leishmania donovani. In humans, the mutation of a GDP-fucose transporter is responsible for an impaired immune response as well as retarded growth. These results suggest that, even though there appear to be a fair number of genes encoding for NSTs, they are not functionally redundant and seem to play specific roles in glycosylation. Correspondence

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Impairment of the Golgi GDP-l-Fucose Transport and Unresponsiveness to Fucose Replacement Therapy in LAD II Patients

Cited by 63 publications

References 35 publications

Differential Terminal Fucosylation of N-Linked Glycans Versus Protein O-Fucosylation in Leukocyte Adhesion Deficiency Type II (CDG IIc)

Differential Terminal Fucosylation of N-Linked Glycans Versus Protein O-Fucosylation in Leukocyte Adhesion Deficiency Type II (CDG IIc)

Golgi nucleotide sugar transport and leukocyte adhesion deficiency II

Nucleotide-sugar transporters: structure, function and roles in vivo

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