The lack of binding of methyl-n-amyl ketone (MAK) to rat liver DNA as demonstrated by direct binding measurements, and 32P-postlabeling techniques.
It has been reported that 14C-labeled methyl-n-amyl ketone (MAK, 2-heptanone) is able to bind spontaneously, in vitro, to isolated rat liver DNA to the extent of 400 pmol/mg DNA; and that 14C-MAK, when given by gavage to female Fischer 344 rats, resulted in HPLC chromatograms of isolated, hydrolyzed liver DNA in which some radiolabel was not associated with the four normal DNA bases dA, dT, dC, and dG. The present studies were undertaken to re-examine the hypothesis that MAK is able to bind to rat liver DNA. In the in vitro study, liver nuclear DNA was incubated with [2-14C]-labeled MAK (25 mCi/mmol) in the absence, or in the presence of rat liver microsomes, precipitated, washed free of unbound MAK, and counted by scintillation spectrometry. No binding to DNA by MAK was detectable. In the in vivo study, groups of five female F344 rats were exposed by inhalation to 0, 80, 400, or 1000 ppm MAK for 6 h/day for 10 days. DNA was purified from the liver nuclei of the 0 and 1000 ppm dosed animals, and 32P-postlabeling techniques were used to assay for adducts. No DNA adducts were detected using these techniques. It was concluded that MAK lacks the ability to bind to rat liver DNA in vitro and in vivo.
Barber
E D
ED
Health and Environment Laboratories, Building 320, Eastman Kodak, Kodak Park, Rochester, NY 14652-6272, USA. ebarber@kodak.com
Miller
K R
KR
Banton
M I
MI
Vijayaraj Reddy
M
M
D000818:Animals; D001345:Autoradiography; D002855:Chromatography, Thin Layer; D004247:DNA / Q000302:isolation & purification; D018736:DNA Adducts / Q000378:metabolism*; D005260:Female; D007659:Ketones / Q000378:metabolism*; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008862:Microsomes, Liver / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D051381:Rats; D011916:Rats, Inbred F344
D018736:DNA Adducts; D007659:Ketones; D010761:Phosphorus Radioisotopes; C011917:2-heptanone; D004247:DNA