Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/7037
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Type: Journal article
Title: Heparan N-sulfatase gene: two novel mutations and transient expression of 15 defects
Author: Esposito, S.
Balzano, N.
Daniele, A.
Villani, G.
Perkins, K.
Weber, B.
Hopwood, J.
Di Natale, P.
Citation: BBA: Molecular Basis of Disease, 2000; 1501(1):1-11
Publisher: Elsevier Science BV
Issue Date: 2000
ISSN: 0925-4439
1878-2434
Statement of
Responsibility: 
Sabrina Esposito, Nicola Balzano, Aurora Daniele, Guglielmo R. D. Villani, Kelly Perkins, Birgit Weber, John J. Hopwood and Paola Di Natale
Abstract: Sanfilippo 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.
Keywords: COS Cells
Animals
Humans
Mucopolysaccharidosis III
Sulfatases
DNA, Complementary
Fluorescent Antibody Technique, Indirect
Blotting, Western
Transfection
Mutagenesis, Site-Directed
Reverse Transcriptase Polymerase Chain Reaction
Gene Expression
Binding Sites
Mutation
Polymorphism, Genetic
Italy
Rights: Copyright © 2000 Published by Elsevier Science B.V.
DOI: 10.1016/S0925-4439(99)00118-0
Appears in Collections:Aurora harvest
Paediatrics publications

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