Sanfilippo D syndrome: estimation of N-acetylglucosamine-6-sulfatase activity with a radiolabeled monosulfated disaccharide substrate.
Sanfilippo D syndrome: estimation of N-acetylglucosamine-6-sulfatase activity with a radiolabeled monosulfated disaccharide substrate.
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Sanfilippo D 综合征：用放射性标记的单硫酸化二糖底物评估 N-乙酰氨基葡萄糖-6-硫酸酯酶活性。
DOI：
10.1016/0003-2697(89)90303-5
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发表时间：
1989
影响因子：
2.9
通讯作者：
J. Hopwood
中科院分区：
文献类型：
--
作者：
C. Freeman;J. Hopwood
N-乙酰葡糖胺-6-硫酸酯酶活性通过孵育放射性标记的二糖O-（α-N-乙酰葡糖胺-6-硫酸盐）-（1 → 3）-l-[6- 3 H]-艾杜糖酸（GlcNAc 6S-IdOA），白细胞匀浆、培养的成纤维细胞和来自正常个体、患有N-乙酰葡糖胺-6-硫酸酯酶缺乏症的患者的尿液（Sanfilippo D综合征，粘多糖沉积症III型）和患有其他粘多糖沉积症和溶酶体贮积症的患者。该测定清楚地区分受影响的纯合子与其专性杂合子和正常对照和其他溶酶体贮积症。成纤维细胞、白细胞和尿液中的硫酸酯酶活性对GlcNAc 6S-IdOA的最适pH分别为4.2、4.5和5.1。成纤维细胞中硫酸酯酶活性的表观Km值为7.2 μm，硫酸根和磷酸根离子均显著抑制硫酸酯酶活性。建议将成纤维细胞或白细胞N-乙酰葡糖胺-6-硫酸酯酶活性对GlcNAc 6S-IdOA的作用用于粘多糖沉积症IIID型患者的常规酶检测和分类。
N-Acetylglucosamine-6-sulfatase activity was assayed by incubation of the radiolabeled disaccharide O-(a-N-acetylglucosamine-6-sulfate)-(1 → 3)-l-[6-3H]-idonic acid (GlcNAc6S-IdOA), with homogenates of leucocytes, cultured fibroblasts, and urine from normal individuals, patients affected with N-acetylglucosamine-6-sulfatase-deficiency (Sanfilippo D syndrome, mucopolysaccharidosis type IIID), and patients affected with other mucopolysaccharidoses and lysosomal storage disorders. The assay clearly distinguished affected homozygotes from their obligate heterozygotes and normal controls and other lysosomal storage disorders. Sulfatase activity in fibroblasts, leucocytes, and urine toward GlcNAc6S-IdOA exhibited a pH optimum at 4.2, 4.5, and 5.1, respectively. Sulfatase activity in fibroblasts had an apparent Kmof 7.2 μm and was significantly inhibited by both sulfate and phosphate ions. The action of fibroblast or leucocyte N-acetylglucosamine-6-sulfatase activity toward GlcNAc6S-IdOA is recommended for the routine enzymatic detection and classification of mucopolysaccharidosis type IIID patients.