Muller V J, Hopwood J J
Clin Genet. 1984 Nov;26(5):414-21. doi: 10.1111/j.1399-0004.1984.tb01081.x.
alpha-L-Iduronidase activity was assayed by incubation of a radiolabelled disaccharide, O-(alpha-L-idopyranosyluronic acid)-(1----3)-2,5 anhydro-D-[1,3H]-talitol 4-sulfate (IdoA-anT4S) derived from dermatan sulfate, with homogenates of leucocytes, cultured amniotic cells and skin fibroblasts from normal individuals and patients affected with an alpha-L-iduronidase-deficiency disorder (mucopolysaccharidosis type I, MPS I), parents of such patients and patients affected with other mucopolysaccharidoses. The assay clearly distinguished affected homozygotes from normal controls, heterozygotes and other mucopolysaccharidosis types. Preliminary results show that fibroblast homogenates from patients with the MPS I Hurler phenotype were virtually unable to hydrolyse IdoA-anT4S, whereas fibroblast homogenates from a patient with a relatively mild (Scheie) phenotype exhibited a residual activity with Vmax value of 2.5 pmol/min/mg protein and an apparent Km of 21 mumol/l compared to a range of 1020-2105 pmol/min/mg for Vmax and 12-35 mumol/l for Km for fibroblasts from normal controls.
通过将一种源自硫酸皮肤素的放射性标记二糖O-(α-L-艾杜糖醛酸)-(1→3)-2,5-脱水-D-[1,3H]-木糖醇4-硫酸盐(艾杜糖醛酸-anhydro-T4S)与正常个体、患有α-L-艾杜糖醛酸缺乏症(I型黏多糖贮积症,MPS I)的患者、此类患者的父母以及患有其他黏多糖贮积症的患者的白细胞、培养的羊膜细胞和皮肤成纤维细胞匀浆一起孵育,来测定α-L-艾杜糖醛酸酶活性。该测定法能明确区分患病纯合子与正常对照、杂合子以及其他类型的黏多糖贮积症。初步结果表明,患有MPS I型Hurler表型的患者的成纤维细胞匀浆几乎无法水解艾杜糖醛酸-anhydro-T4S,而患有相对轻度(Scheie)表型的患者的成纤维细胞匀浆表现出残余活性,其Vmax值为2.5 pmol/分钟/毫克蛋白质,表观Km为21 μmol/升,相比之下,正常对照的成纤维细胞的Vmax范围为1020 - 2105 pmol/分钟/毫克,Km为12 - 35 μmol/升。