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肠道糖苷酶的免疫组织化学定位

Immunohistochemical localization of intestinal glycosidases.

作者信息

Slabý J, Lojda Z, Kraml J, Kolínská J

出版信息

Acta Univ Carol Med Monogr. 1977(77 Pt 1):105-11.

PMID:97928
Abstract

The presence of antigenic determinants of the following enzymes was detected in enterocytes by the indirect immunofluorescence method: 1. lactase in human biopsy material, 2. sucrase-isomaltase during ontogenesis in the rat. 1. Lactase: The antigenic relationship between rat and human lactase, demonstrated with the isolated enzymes, was utilized for the histochemical localization of human lactase. The indirect immunofluorescence method, using guinea pig antiserum to rat lactase, demonstrated the presence of human lactase in the enterocyte brush border. The usefulness of this method for clinical practice resides in the possibility of detecting enzymatically inactive protein immunologically related to lactase in cases of lactase deficiency, thereby facilitating more detailed classification of these diseases. 2. Sucrase-isomaltase: Guinea pig antiserum to rat sucrase-isomaltase (SI) was prepared. It was used to demonstrate antigenic determinants of the enzyme in the enterocyte brush border of the rat during ontogenesis. Structural SI protein is already present in 3-day-old rats, whereas enzyme activity can first be demonstrated histochemically from the 11th day of life and biochemically, in vitro, not until about the 18th day. We consider that this technique can be used for studying the biogenesis of membrane-bound enzymes.

摘要

采用间接免疫荧光法在肠细胞中检测到了以下几种酶的抗原决定簇

  1. 人体活检材料中的乳糖酶;2. 大鼠个体发育过程中的蔗糖酶 - 异麦芽糖酶。1. 乳糖酶:利用分离出的大鼠和人乳糖酶之间的抗原关系,对人乳糖酶进行组织化学定位。采用豚鼠抗大鼠乳糖酶血清的间接免疫荧光法,证实了人乳糖酶存在于肠细胞刷状缘。该方法在临床实践中的用途在于,在乳糖酶缺乏的情况下,有可能检测到与乳糖酶免疫相关的无酶活性蛋白,从而有助于对这些疾病进行更详细的分类。2. 蔗糖酶 - 异麦芽糖酶:制备了豚鼠抗大鼠蔗糖酶 - 异麦芽糖酶(SI)血清。用它来证实大鼠个体发育过程中肠细胞刷状缘该酶的抗原决定簇。结构型SI蛋白在3日龄大鼠中就已存在,而酶活性直到出生后第11天才能通过组织化学方法证实,在体外生化检测中直到约第18天才能检测到。我们认为该技术可用于研究膜结合酶的生物发生。

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