Heparan N-sulfatase gene: two novel mutations and transient expression of 15 defects

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dc.contributor.authorEsposito, S.
dc.contributor.authorBalzano, N.
dc.contributor.authorDaniele, A.
dc.contributor.authorVillani, G.
dc.contributor.authorPerkins, K.
dc.contributor.authorWeber, B.
dc.contributor.authorHopwood, J.
dc.contributor.authorDi Natale, P.
dc.date.issued2000
dc.description.abstractSanfilippo syndrome type A or mucopolysaccharidosis IIIA (MPS IIIA) results from the deficiency of the enzyme heparan N-sulfatase (NS, EC 3.10.1.1), required for the degradation of heparan sulfate. Molecular defects of 24 Italian MPS IIIA patients were recently reported by our group. We report here two novel mutations: 1040insT and Q365X and the expression studies on 15 of the identified defects. Transient expression of COS cells by cDNA mutagenized to correspond to heparan N-sulfatase mutations Y40N, A44T, 166delG, G122R, P128L, L146P, R150Q, D179N, R182C, R206P, P227R, 1040insT, 1093insG, E369K, R377C did not yield active enzyme, demonstrating the deleterious nature of the mutations. Western blot analysis and metabolic labeling experiments revealed, for cells transfected with wild-type enzyme, a precursor 62-kDa form and a mature 56-kDa form. Western blot resulted, for 11 mutations, in the presence of both forms, indicating a normal maturation of the mutant enzyme. Western blot, metabolic labeling and immunofluorescence experiments suggested, for mutations 166delG, L146P, 1040insT and 1093insG, an increased degradation of the mutant enzymes.
dc.description.statementofresponsibilitySabrina Esposito, Nicola Balzano, Aurora Daniele, Guglielmo R. D. Villani, Kelly Perkins, Birgit Weber, John J. Hopwood and Paola Di Natale
dc.identifier.citationBiochimica et Biophysica Acta - Molecular Basis of Disease, 2000; 1501(1):1-11
dc.identifier.doi10.1016/S0925-4439(99)00118-0
dc.identifier.issn0925-4439
dc.identifier.issn1878-2434
dc.identifier.urihttp://hdl.handle.net/2440/7037
dc.language.isoen
dc.publisherElsevier Science BV
dc.rightsCopyright © 2000 Published by Elsevier Science B.V.
dc.source.urihttps://doi.org/10.1016/s0925-4439(99)00118-0
dc.subjectCOS Cells
dc.subjectAnimals
dc.subjectHumans
dc.subjectMucopolysaccharidosis III
dc.subjectSulfatases
dc.subjectDNA, Complementary
dc.subjectFluorescent Antibody Technique, Indirect
dc.subjectBlotting, Western
dc.subjectTransfection
dc.subjectMutagenesis, Site-Directed
dc.subjectReverse Transcriptase Polymerase Chain Reaction
dc.subjectGene Expression
dc.subjectBinding Sites
dc.subjectMutation
dc.subjectPolymorphism, Genetic
dc.subjectItaly
dc.titleHeparan N-sulfatase gene: two novel mutations and transient expression of 15 defects
dc.typeJournal article
pubs.publication-statusPublished

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