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Title

Molecular and functional characterization of a new X-linked chronic granulomatous disease variant (X91+) case with a double missense mutation in the cytosolic gp91phox C-terminal tail

Authors
Stasia, Marie José
Rousseau, Pascale
Martel, Cécile
Bordigoni, Pierre
Published in Biochimica et Biophysica Acta. 2002, vol. 1586, no. 3, p. 316-30
Abstract We report here two atypical cases of X-linked CGD patients (first cousins) in which cytochrome b(558) is present at a normal level but is not functional (X91+). The mutations were localized by single-strand conformational polymorphism of reverse transcriptase-polymerase chain reaction amplified fragments and then identified by sequence analysis. They consisted in two base substitutions (C919 to A and C923 to G), changing His303 to Asn and Pro304 to Arg in the cytosolic gp91phox C-terminal tail. Mismatched polymerase chain reaction and genomic DNA sequencing showed that mothers had both wild-type and mutated alleles, confirming that this case was transmitted in an X-linked fashion. A normal amount of FAD was found in neutrophil membranes, both in the X91+ patients and their parents. Epstein-Barr virus-transformed B lymphocytes from the X91+ patients acidified normally upon stimulation with arachidonic acid, indicating that the mutated gp91phox still functioned as a proton channel. A cell-free translocation assay demonstrated that the association of the cytosolic factors p47phox and p67phox with the membrane fraction was strongly disrupted. We concluded that residues 303 and 304 are crucial for the stable assembly of the NADPH oxidase complex and for electron transfer, but not for its proton channel activity.
Keywords Cell Membrane/metabolismCytochrome b Group/metabolismCytosol/metabolismFlavin-Adenine Dinucleotide/analysisGranulomatous Disease, Chronic/blood/genetics/metabolismHumansInfantMaleMembrane Glycoproteins/genetics/metabolismMutation, MissenseN-Formylmethionine Leucyl-PhenylalanineNADPH Oxidase/metabolismNeutrophils/enzymologyPolymorphism, Single-Stranded ConformationalRNA, Messenger/metabolismTetradecanoylphorbol Acetate
Identifiers
PMID: 11997083
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Research group Signaux intracellulaires (210)
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STASIA, Marie José et al. Molecular and functional characterization of a new X-linked chronic granulomatous disease variant (X91+) case with a double missense mutation in the cytosolic gp91phox C-terminal tail. In: Biochimica et Biophysica Acta, 2002, vol. 1586, n° 3, p. 316-30. https://archive-ouverte.unige.ch/unige:30346

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Deposited on : 2013-10-11

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