Diagnosis of adult gaucher disease: Use of a new chromogenic substrate, 2-hexadecanoylamino-4-nitrophenyl-β-d-glucopyranoside, in cultured skin fibroblasts

TY - JOUR

T1 - Diagnosis of adult gaucher disease

T2 - Use of a new chromogenic substrate, 2-hexadecanoylamino-4-nitrophenyl-β-d-glucopyranoside, in cultured skin fibroblasts

AU - Johnson, William G.

AU - Gal, Andrew E.

AU - Miranda, Armand F.

AU - Pentchev, Peter G.

N1 - Funding Information: This work was supported by grants from The National Institutes of Health (No. l-ROl-NS-15281), the National Foundation, the Muscular Dystrophy Association, Inc. (H. Houston Merritt Clinical Center for Muscular Dystrophy and Related Diseases), and by a Career Scientist Award from the Irma T. Hirsch1 Foundation. The authors wish to thank David Heinricher, Cheryl Cohen and Jane Fouks for expert technical assistance.

PY - 1980/3/14

Y1 - 1980/3/14

N2 - 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-β-d-glucopyranoside are commonly used for its diagnosis. We studied the use of a new chromogenic substrate, 2-hexadecanoylamino-4-nitrophenyl-β-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 fibro blast protein added up to 150 μg per incubation. The pH optimum was 4.8. The Km was 0.19 mmol/1. 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.

AB - 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-β-d-glucopyranoside are commonly used for its diagnosis. We studied the use of a new chromogenic substrate, 2-hexadecanoylamino-4-nitrophenyl-β-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 fibro blast protein added up to 150 μg per incubation. The pH optimum was 4.8. The Km was 0.19 mmol/1. 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.

UR - https://www.scopus.com/pages/publications/0018837396

UR - https://www.scopus.com/pages/publications/0018837396#tab=citedBy

U2 - 10.1016/0009-8981(80)90437-4

DO - 10.1016/0009-8981(80)90437-4

M3 - Article

C2 - 7389109

SN - 0009-8981

VL - 102

SP - 91

EP - 97

JO - Clinica Chimica Acta

JF - Clinica Chimica Acta

IS - 1

ER -