Gualandi S, Govoni M, Ricciardelli P, Rossini R, Tazzari R
Boll Soc Ital Biol Sper. 1984 May 30;60(5):937-43.
We examined the activity of total N-acetyl-beta-hexosaminidase and of its isoenzyme forms, that represent different stages of the maturation of the lysosomal hydrolase. In both methods the enzyme catalyzes the separation of 4-methylumbelliferone, a fluorescent substance, from 4-methylumelliferyl-2-acetamido-2-deoxy-beta-D-glucopyranoside. We used Leaback's method for the fluorimetric assay of total enzyme, and Ellis's DEAE-cellulose microcolum chromatography for the assay of its components. We obtained a clear separation of each fraction. We will apply these methods in our further studies of children with renal damage, because hexosaminidase seems to be one of the most sensitive markers of tubular damage.
我们检测了总N-乙酰-β-己糖胺酶及其同工酶形式的活性，这些同工酶代表了溶酶体水解酶成熟的不同阶段。在这两种方法中，该酶催化荧光物质4-甲基伞形酮从4-甲基伞形酮基-2-乙酰氨基-2-脱氧-β-D-吡喃葡萄糖苷中分离出来。我们使用Leaback法进行总酶的荧光测定，并使用Ellis的DEAE-纤维素微柱色谱法测定其成分。我们得到了每个组分的清晰分离。我们将把这些方法应用于我们对肾损伤儿童的进一步研究中，因为己糖胺酶似乎是肾小管损伤最敏感的标志物之一。
