Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/7708
Citations | ||
Scopus | Web of ScienceĀ® | Altmetric |
---|---|---|
?
|
?
|
Type: | Journal article |
Title: | Expression and functional characterization of human mutant sulfamidase in insect cells |
Author: | Montfort, M. Garrido, E. Hopwood, J. Grinberg, D. Chabas, A. Vilageliu, L. |
Citation: | Molecular Genetics and Metabolism, 2004; 83(3):246-251 |
Publisher: | Academic Press Inc Elsevier Science |
Issue Date: | 2004 |
ISSN: | 1096-7192 1096-7206 |
Abstract: | Mucopolysaccharidosis IIIA (MPS IIIA; Sanfilippo syndrome) is an autosomal recessive lysosomal disorder caused by the deficiency of sulfamidase (EC 3.10.1.1), required for the degradation of the mucopolysaccharide heparan sulfate. The molecular defects of 26 unrelated Spanish MPS IIIA patients were recently reported by our group. Here we describe the heterologous expression, using a baculovirus system, of the cDNAs corresponding to eight out of the 14 mutant alleles present in this patient group and the characterization of the corresponding mutant enzymes. In particular, we expressed the following alleles: p.S66W, p.R74H, p.Q85R, p.R206P, p.L386R, p.R433W, p.R433Q, and c.1079delC (previously named as c.1091delC), and the two variants of the polymorphism p.R456H. The expression of the mutant alleles and the characterization of the corresponding enzymes revealed that their activity was severely compromised. Only mutations p.S66W and p.R206P retained low levels of residual activity. However, Western blot analysis showed in all cases the presence of the expected two forms of the sulfamidase, the precursor and the mature proteins, indicating a normal processing of the mutant enzyme. |
Keywords: | Cells, Cultured Animals Humans Spodoptera Mucopolysaccharidosis III Hydrolases DNA, Complementary DNA Primers Blotting, Western Electrophoresis, Polyacrylamide Gel Mutagenesis, Site-Directed Gene Expression Genotype Mutation Alleles Spain |
DOI: | 10.1016/j.ymgme.2004.07.001 |
Published version: | http://dx.doi.org/10.1016/j.ymgme.2004.07.001 |
Appears in Collections: | Aurora harvest Paediatrics publications |
Files in This Item:
There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.