Abstract |
The biochemical properties of hexosaminidase A (HexA) and the coding sequence of the alpha-subunit were examined in a patient of Syrian ancestry with the B1 form of Tay-Sachs disease (TSD). The biochemical characteristics of the variant HexA suggest that both active sites are affected by the mutation(s). Kinetic studies with the beta-subunit specific substrate, 4-methylumbelliferyl-beta-D-N-acetylglucosamine (MUG), revealed a significant difference between the Km values. of normal and variant HexA, while no difference was found when the sulfated substrate MUG-6-sulfate (MUGS), which is specific for the alpha-subunit active site, was used. The Vmax values for both substrates were significantly lower in extracts from B1 variant cells than in control extracts, implying a reduced enzyme level in the variant cells. A noncompetitive inhibitor of the reaction with MUGS, N-acetylglucosamine (NAG), induced a significant inhibition (30%) in the mutant cells only. When MUG was used as substrate, variant HexA was found to be more heat stable (T50 = 170 min) than normal HexA (T50 = 65 min). Furthermore, the mutant cell preparation differed from control in the relation between Hex thermosensitivity and protein concentration in the reaction. Two new mutations were identified in exon 5 of the HexA gene: a C496 to G transversion, which produced an Arg166 -->Gly alteration and a deletion of C498 which generated a shift in the reading frame. The patient was a heterozygote for both mutations even though her parents are first cousins. There is no evidence as yet which of these mutations accounts for the B1 phenotype.
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Authors | L Peleg, F Meltzer, M Karpati, B Goldman |
Journal | Biochemical and molecular medicine
(Biochem Mol Med)
Vol. 54
Issue 2
Pg. 126-32
(Apr 1995)
ISSN: 1077-3150 [Print] United States |
PMID | 8581357
(Publication Type: Journal Article)
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Chemical References |
- Hexosaminidase A
- beta-N-Acetylhexosaminidases
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Topics |
- Base Sequence
- Cells, Cultured
- Exons
- Female
- Fibroblasts
(cytology, enzymology)
- Hexosaminidase A
- Humans
- Kinetics
- Male
- Molecular Sequence Data
- Mutation
- Polymorphism, Single-Stranded Conformational
- Tay-Sachs Disease
(enzymology, etiology, genetics)
- beta-N-Acetylhexosaminidases
(antagonists & inhibitors, chemistry, physiology)
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