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使用凝集素作为探针标记细胞表面碳水化合物的定性和定量方面。

Qualitative and quantitative aspects of labeling cell surface carbohydrates using lectins as probes.

作者信息

Lotan R

出版信息

Scan Electron Microsc. 1979(3):549-64.

PMID:392714
Abstract

Lectins are proteins which bind mono- and oligosaccharides with great specificity. Many polysaccharides, glycoproteins and glycolipids which are important constituents of cell walls and surface membranes of prokaryotic and eukaryotic cells, contain sugar moieties with which lectins can interact. As a result lectins have been extensively used for the study of cell surface and membrane structure of their labeling. This paper reviews and evaluates the available methods for the preparation of fluorescent, electron-dense and radioactive lectin derivatives. The procedures involved in the visualization of lectin binding under light and electron microscopy are described. In addition, the methods for quantitative evaluation of the fluorescence on labeled cells and for analysis of the number of cell surface lectin binding sites using radioactively-labeled lectins are outlined. Examples are given for the application of fluorescent lectin derivatives for the detection of specific saccharide-containing molecules on the surfaces of living or fixed microbial cells and various normal and neoplastic cells. The use of lectins to demonstrate the dynamic nature of cell membranes and to detect changes in membrane structure or organization which occur or organization which occur during differentiation, development or after neoplastic transformation is discussed.

摘要

凝集素是一类能高度特异性结合单糖和寡糖的蛋白质。许多多糖、糖蛋白和糖脂是原核细胞和真核细胞细胞壁及表面膜的重要组成成分,它们含有能与凝集素相互作用的糖基。因此,凝集素已被广泛用于研究细胞表面和膜结构及其标记。本文综述并评估了制备荧光、电子致密和放射性凝集素衍生物的现有方法。描述了在光学显微镜和电子显微镜下观察凝集素结合的相关步骤。此外,还概述了定量评估标记细胞上荧光的方法以及使用放射性标记凝集素分析细胞表面凝集素结合位点数量的方法。文中给出了荧光凝集素衍生物用于检测活的或固定的微生物细胞以及各种正常和肿瘤细胞表面特定含糖类分子的应用实例。还讨论了利用凝集素来证明细胞膜的动态性质以及检测在分化、发育过程中或肿瘤转化后发生的膜结构或组织变化。

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