Diagnosis of adult Gaucher disease: use of a new chromogenic substrate, 2-hexadecanoylamino-4-nitrophenyl-beta-D-glucopyranoside, in cultured skin fibroblasts.
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| Abstract |
Gaucher disease is a group of lipid storage diseases in which the glycosphingolipid glucocerebroside accumulates in tissues because of deficiency of the enzyme glucocerebrosidase. Radioactively labelled glucocerebroside and the artificial fluorogenic substrate 4-methylumbelliferyl-beta-D-glucopyranoside are commonly used for its diagnosis. We studied the use of a new chromogenic substrate, 2-hexadecanoylamino-4-nitrophenyl-beta-D-glucopyranoside in cultured skin fibroblasts. The amount of reaction product, 2-hexadecanoylamino-4-nitrophenol, increased linearly with incubation time for at least 4 h and was proportional to the amount of fibroblast protein added up to 150 micrograms per incubation. The pH optimum was 4.8. The Km was 0.19 mmol/l. The mean activity of control cultured skin fibroblasts was 22.9 +/- 5.4 nmol of product formed per mg fibroblast protein per hour under standard conditions. Cultured skin fibroblasts from patients with adult non-neuropathic Gaucher disease had reduced activity, 6.4 +/- 2.4 nmol/mg/h or 28% of control activity. This compared well with mean enzyme activity in the same patients determined using the natural substrate, [14C]glucocerebroside: 28% of control activity. Heterozygotes had reduced activity with the new substrate.
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| Authors | W G Johnson, A E Gal, A F Miranda, P G Pentchev |
| Journal | Clinica chimica acta; international journal of clinical chemistry (Clin Chim Acta) Vol. 102 Issue 1 Pg. 91-7 (Mar 14 1980) ISSN: 0009-8981 [Print] Netherlands |
| PMID | 7389109 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.) |
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