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A simple radioassay of benzo(a)pyrene activation: Observations on the covalent interactions of benzo(a)pyrene with protein

✍ Scribed by Forrest B. Thomas; N.Burr Furlong

Publisher: Elsevier Science
Year: 1976
Tongue: English
Weight: 444 KB
Volume: 72
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(76)90565-0

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✦ Synopsis

A simple radioassay method is described to measure specifically and quantitatively the metabolic activation of the carcinogen benzo(a)pyrene (BP) to a derivative which reacts covalently with a macromolecular target molecule (protein). The NADPH*-dependent formation of this ma~romoiecule-hydrocarbon complex during incubation with rat liver microsomes is linear over a 30 min period, and occurs at a rate which is approximately 60% that of phenolic BP derivatives. No such product is formed during incubation with either heatinactivated microsome preparations or bovine serum albumin. inhibition of this reaction by sodium azide is seen only at concentrations of 0.1 M or greater, and is not distinguishable from inhibition by sodium chloride at similar levels. These observations suggest that Fe3+-proto~~hyrin is not essential for BP activation.

Methods

Benzo(a)pyrene was obtained from Aldrich Chemical Company, and is used in this laboratory as a 2 mM solution in dimethylsulfoxide (DMSO).

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