Kekic M, dos Remedios C G
Department of Anatomy and Histology, Institute for Biomedical Research, The University of Sydney, Australia.
Electrophoresis. 1999 Jul;20(10):2053-8. doi: 10.1002/(SICI)1522-2683(19990701)20:10<2053::AID-ELPS2053>3.0.CO;2-S.
We describe how the interaction between actin and its protein ligands can be used to evaluate the presence of certain metal (Cd, Cu, Hg, Zn) ions and organic compounds (2,4-dioxin or Picloram) which are common components of environmental pollution. The assay detects the high-affinity binding of actin to actin-binding proteins (ABPs), cofilin or DNase I. The actin-ABP complex was analyzed using native polyacrylamide gel electrophoresis and quantified by scanning densitometry. These proteins are widely distributed in animals and plant cells. The assay involves allowing the proteins to form an actin-ABP complex into which increasing amounts of pollutants are titrated. Thus, the assay directly tests for inhibition of protein-protein interaction. It is sensitive to common pollutants using concentration ranges over which they are known to exert a biological toxicity. A convenient feature of the assay is the fact that all the proteins can be stored in freeze-dried form, and can be purchased commercially. We suggest that if this molecular assay is sensitive to a wide range of environmental pollutants, it could be used as a rapid and convenient assay of the environment in combination with currently available tests.
我们描述了肌动蛋白与其蛋白质配体之间的相互作用如何用于评估某些金属（镉、铜、汞、锌）离子和有机化合物（2,4-二恶英或毒莠定）的存在，这些是环境污染的常见成分。该检测方法可检测肌动蛋白与肌动蛋白结合蛋白（ABP）、丝切蛋白或脱氧核糖核酸酶I的高亲和力结合。使用天然聚丙烯酰胺凝胶电泳分析肌动蛋白-ABP复合物，并通过扫描密度计进行定量。这些蛋白质广泛分布于动物和植物细胞中。该检测方法包括让蛋白质形成肌动蛋白-ABP复合物，然后向其中滴定越来越多的污染物。因此，该检测方法直接测试蛋白质-蛋白质相互作用的抑制情况。它对常见污染物敏感，所使用的浓度范围是已知它们会产生生物毒性的范围。该检测方法的一个便利之处在于，所有蛋白质都可以以冻干形式储存，并且可以从商业渠道购买。我们认为，如果这种分子检测方法对多种环境污染物敏感，那么它可以与目前可用的检测方法结合，用作一种快速便捷的环境检测方法。
