用伴刀豆球蛋白A-琼脂糖亲和色谱法测定人脑脊液的酸性糖苷酶和芳基硫酸酯酶活性。

Acid glycosidase and arylsulfatase activities of human cerebrospinal fluid as measured by concanavalin A-sepharose affinity chromatography.

作者信息

Cherian R, Balasubramanian A S

出版信息

Clin Chim Acta. 1978 Nov 1;89(3):411-6. doi: 10.1016/0009-8981(78)90403-5.

Abstract

An affinity chromatographic method using concanavalin A-Sepharose is described for the determination of N-acetyl-beta-D-glucosaminidase, arylsulfatase. alpha-L-Fucosidase and alpha-D-mannosidase activities in the human cerebrospinal fluid. By this method (starting with 12 to 20 ml samples of cerebrospinal fluid) the above enzymes could be obtained in a concentrated form and their activities could be determined within incubation periods of 30 min to 1 h under the assay conditions described. The pH optima of the enzymes were in the range of pH 4 to 5. About 80% of the total cerebrospinal fluid N-acetyl-beta-D-glucosaminidase was found to be the A form by DEAE-Sephadex A-50 chromatography. About 60% of the total arylsulfatase was also found to be the A form. Determination of these enzyme activities in a few samples of human cerebrospinal fluid indicated a rough proportionality between the enzyme activities and the protein concentration in the cerebrospinal fluid.

摘要

描述了一种使用伴刀豆球蛋白A-琼脂糖的亲和色谱法，用于测定人脑脊液中的N-乙酰-β-D-氨基葡萄糖苷酶、芳基硫酸酯酶、α-L-岩藻糖苷酶和α-D-甘露糖苷酶的活性。通过这种方法（从12至20毫升脑脊液样品开始），上述酶可以以浓缩形式获得，并且在所述测定条件下，其活性可以在30分钟至1小时的温育期内测定。这些酶的最适pH在pH 4至5范围内。通过DEAE-葡聚糖A-50色谱法发现，脑脊液中约80%的总N-乙酰-β-D-氨基葡萄糖苷酶为A型。还发现约60%的总芳基硫酸酯酶也是A型。对几份人脑脊液样品中这些酶活性的测定表明，酶活性与脑脊液中的蛋白质浓度之间大致成比例。