Immunoassays for proteins alkylated by nicotine-derived N-nitrosamines.
Polyclonal antibodies recognizing the pyridyloxobutyl (POB) moiety of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) were produced in rabbits immunized either with POB-bovine albumin or POB-Sepharose. The POB intermediates necessary to modify the protein were generated by alkaline (pH 9.0) treatment of the synthetic precursor 4-(carbethoxynitrosamino)-1-(3-pyridyl)-1-butanone. In a competitive enzyme linked immunoabsorbent assay (ELISA), 70 pmole NNK inhibited 50% of the binding of the anti-POB antibodies to POB-protein absorbed on microtiterplates. This 50% inhibition varied from 70 pmole to 200 nmole using a series of NNK analogues, depending on the integrity of the POB moiety. Immunological techniques initiated in this study detect NNK-protein conjugates or measure the quantity of POB groups liberated upon alkaline or acid treatment of NNK modified protein.
Talbot
B
B
Department of Biology, Faculty of Sciences, University of Sherbrooke, Québec, Canada.
Desnoyers
S
S
Castonguay
A
A
D000818:Animals; D002273:Carcinogens / Q000633:toxicity*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006868:Hydrolysis; D007118:Immunoassay; D015151:Immunoblotting; D007525:Isoelectric Focusing; D007694:Killer Cells, Natural / Q000187:drug effects; D009538:Nicotine / Q000031:analogs & derivatives*; D009602:Nitrosamines / Q000633:toxicity*; D011506:Proteins / Q000032:analysis*
D002273:Carcinogens; D009602:Nitrosamines; D011506:Proteins; D009538:Nicotine; C016583:4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone; C008655:N'-nitrosonornicotine