Electrophoretic monitoring of pollutants: effect of cations and organic compounds on protein interactions monitored by native gel electrophoresis.
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.
Kekic
M
M
Department of Anatomy and Histology, Institute for Biomedical Research, The University of Sydney, Australia.
dos Remedios
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CG
D051339:Actin Depolymerizing Factors; D000199:Actins / Q000378:metabolism; D000818:Animals; D002104:Cadmium / Q000494:pharmacology; D002412:Cations / Q000494:pharmacology*; D002645:Chickens; D003300:Copper / Q000494:pharmacology; D003850:Deoxyribonuclease I / Q000378:metabolism; D004147:Dioxins / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D008628:Mercury / Q000494:pharmacology; D008840:Microfilament Proteins / Q000378:metabolism; D010846:Picloram / Q000494:pharmacology; D011506:Proteins / Q000378:metabolism*; D011817:Rabbits; D014873:Water Pollutants / Q000032:analysis*; D015032:Zinc / Q000494:pharmacology
D051339:Actin Depolymerizing Factors; D000199:Actins; D002412:Cations; D004147:Dioxins; D008840:Microfilament Proteins; D011506:Proteins; D014873:Water Pollutants; D002104:Cadmium; D003300:Copper; D003850:Deoxyribonuclease I; D008628:Mercury; D015032:Zinc; D010846:Picloram