一种用于诊断A型粘多糖贮积症（MPS IV A）的荧光酶测定法。

A fluorimetric enzyme assay for the diagnosis of Morquio disease type A (MPS IV A).

作者信息

van Diggelen O P, Zhao H, Kleijer W J, Janse H C, Poorthuis B J, van Pelt J, Kamerling J P, Galjaard H

机构信息

Department of Clinical Genetics, University Hospital, Erasmus University, Rotterdam, The Netherlands.

出版信息

Clin Chim Acta. 1990 Feb 28;187(2):131-9. doi: 10.1016/0009-8981(90)90339-t.

DOI:10.1016/0009-8981(90)90339-t

PMID:2107987

Abstract

4-Methylumbelliferyl-beta-D-galactopyranoside-6-sulphate was synthesized and used for the determination of galactose-6-sulphate sulphatase activity. Fibroblasts and leucocytes from 12 different Morquio A patients, showed 0.0-2.7% of mean normal galactose-6-sulphate sulphatase activity. Heterozygotes showed intermediate activities. The enzymatic liberation of the fluorochrome from 4-methylumbelliferyl-beta-D-galactopyranoside-6-sulphate requires the sequential action of galactose-6-sulphate sulphatase and beta-galactosidase. Normal beta-galactosidase activity caused nearly complete hydrolysis of non-fluorescing 4-methylumbelliferyl-galactoside, formed during incubation. In cell extracts with a beta-galactosidase deficiency however, a second incubation in the presence of excess beta-galactosidase is needed to avoid underestimation of galactose-6-sulphate sulphatase activity.

摘要

合成了4-甲基伞形酮基-β-D-吡喃半乳糖苷-6-硫酸盐，并将其用于测定6-硫酸半乳糖硫酸酯酶的活性。来自12名不同的Morquio A患者的成纤维细胞和白细胞显示出平均正常6-硫酸半乳糖硫酸酯酶活性的0.0 - 2.7%。杂合子表现出中等活性。从4-甲基伞形酮基-β-D-吡喃半乳糖苷-6-硫酸盐中酶促释放荧光染料需要6-硫酸半乳糖硫酸酯酶和β-半乳糖苷酶的顺序作用。正常的β-半乳糖苷酶活性会导致在孵育过程中形成的无荧光的4-甲基伞形酮基半乳糖苷几乎完全水解。然而，在β-半乳糖苷酶缺乏的细胞提取物中，需要在过量β-半乳糖苷酶存在下进行第二次孵育，以避免低估6-硫酸半乳糖硫酸酯酶的活性。