放射免疫分析法测定岩藻糖苷贮积症中α-L-岩藻糖苷酶蛋白含量的降低
Radioimmunoassay determination of decreased amounts of alpha-L-fucosidase protein in fucosidosis.
作者信息
Andrews-Smith G L, Alhadeff J A
出版信息
Biochim Biophys Acta. 1982 Mar 15;715(1):90-6. doi: 10.1016/0304-4165(82)90053-8.
Purified human liver alpha-L-fucosidase (EC 3.2.1.51) has been radioiodinated by a chloramine-T procedure to a specific activity of 3.7 . 10(6) dpm/micrograms protein without altering its apparent Michaelis constant for the 4-methylumbelliferyl substrate. This 125I-labeled alpha-L-fucosidase has been used in development of a competitive binding radioimmunoassay for alpha-L-fucosidase which can detect 1-2 ng of enzyme protein and has been employed to quantify the amount of alpha-L-fucosidase protein in the liver and spleen from a patient with fucosidosis. Less than 1% of the normal amount of alpha-L-fucosidase protein is present suggesting that normal amounts of catalytically inactive alpha-L-fucosidase are not found in this disease.
纯化的人肝α-L-岩藻糖苷酶（EC 3.2.1.51）已通过氯胺-T法进行放射性碘化，比活度达到3.7×10⁶ dpm/μg蛋白质，且未改变其对4-甲基伞形酮底物的表观米氏常数。这种¹²⁵I标记的α-L-岩藻糖苷酶已用于开发α-L-岩藻糖苷酶的竞争性结合放射免疫测定法，该方法可检测1-2 ng酶蛋白，并已用于定量岩藻糖苷贮积症患者肝脏和脾脏中α-L-岩藻糖苷酶蛋白的含量。结果表明，该患者体内α-L-岩藻糖苷酶蛋白含量不到正常水平的1%，提示在这种疾病中未发现正常量的催化无活性的α-L-岩藻糖苷酶。
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