A 3H-labelled trisaccharide from heparin as substrate for acetyl:CoA: 2-amino-2-deoxy-α-d-glucoside N-acetyltransferase
✍ Scribed by Udo Klein; Kurt von Figura
- Publisher
- Elsevier Science
- Year
- 1980
- Tongue
- English
- Weight
- 329 KB
- Volume
- 78
- Category
- Article
- ISSN
- 0008-6215
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✦ Synopsis
ABSIRACT
The tetrasaccharide fraction obtained by gel chromatography after treatment of commercially available heparin with nitrous acid was reduced with NaB3H4 and then hydrolysed with 2M trifluoracetic acid at 70 ° for 3 days. By gel chromatography and electrophoresis, the 3H-labelled trisaccharide 1 bearing an unsubstituted 2-amino-2-deoxy-o-glucosyl group in the non-reducing position was obtained (18~o from the 3H-labelled tetrasaccharide). By sequential, enzymic degradation, the structure :~-o-GlcN-(l~4)-fl-D-GlcA-(l~4)-[I-3H]aManol t was obtained for 1, which is a substrate for acetyl-CoA: 2-amino-2-deoxy-~-D-glucoside N-acetyltransferase, an enzyme that is deficient in the Sanfilippo C syndrome. In human-skin fibroblasts, the pH optimum of acetyl transfer onto 1 was between pH 5.5 and 7.0, and dependent on the buffer. An apparent K m for 1 of 0.14mM was found.