Dean M F
J Inherit Metab Dis. 1983;6(3):108-11. doi: 10.1007/BF01800738.
The substrate O-(alpha-L-idopyranosyluronic acid-2-sulphate)-(1 leads to 4)-2,5-anhydro-D-(3H-1) mannitol-6-sulphate was used at a final concentration of 50 mmol/l to measure the alpha-L-idurono-2-sulphate sulphatase activities of cell extracts, serum and amniotic fluid. Activities were measured after dialysis against water, to avoid the inhibitory effect of sodium chloride and the reaction products separated by ion-exchange chromatography on ECTEOLA cellulose. The enzyme present in normal serum had an apparent Km of 0.12 mmol/l. The mean activities of normal serum, fibroblasts and leukocytes were 0.61, 16.63 and 18.75 nmol/mg protein per hour respectively, while corresponding 'Hunter' tissues contained between 1% and 3% of normal activity. Cultured normal amniotic cells had a mean activity of 8.46 nmol/mg protein per hour, while those cultured from 'Hunter' patients contained about 6% of normal activity. Hunter disease can thus be readily diagnosed in a wide variety of tissues. Samples from three obligate heterozygotes were found to have iduronate sulphatase activities amounting to 41%, 57% and 63% of normal values.
以终浓度50 mmol/l使用底物O-(α-L-艾杜吡喃糖醛酸-2-硫酸酯)-(1→4)-2,5-脱水-D-(3H-1)甘露醇-6-硫酸酯来测定细胞提取物、血清和羊水的α-L-艾杜糖醛酸-2-硫酸酯硫酸酯酶活性。在对水进行透析后测量活性，以避免氯化钠的抑制作用以及离子交换色谱法在ECTEOLA纤维素上分离的反应产物的影响。正常血清中存在的酶的表观Km为0.12 mmol/l。正常血清、成纤维细胞和白细胞的平均活性分别为每小时0.61、16.63和18.75 nmol/mg蛋白质，而相应的“亨特”组织的活性为正常活性的1%至3%。培养的正常羊膜细胞的平均活性为每小时8.46 nmol/mg蛋白质，而从“亨特”患者培养的细胞的活性约为正常活性的6%。因此，在多种组织中都可以很容易地诊断出亨特病。发现来自三名 obligate杂合子的样本的艾杜糖醛酸硫酸酯酶活性分别为正常值的41%、57%和63%。
