DNA-protein crosslinking by trans-platinum(II)diamminedichloride in mammalian cells, a new method of analysis.
DNA-protien crosslinks produced in mouse leukemia L1210 cells by trans-Pt(II)diamminedichloride were quantitated using the technique of DNA alkaline elution. DNA single-strand segments that were or were not linked to protein were separable into distinct components by alkaline elution after exposure of the cells to 2--15 kR of X-ray. Protein-linked DNA strands were separated on the basis of their retention of filters at pH 12 while free DNA strands of the size generated by 2--15 kR of X-ray passed rapidly through the filters. The retention of protein-linked DNA strands was attributable to adsorption of protein to the filter under the conditions of alkaline elution. The results obeyed a simple quantitative model according to which the frequency of DNA-protein crosslinks could be calculated.
D000818:Animals; D002460:Cell Line; D002945:Cisplatin / Q000494:pharmacology*; D004273:DNA, Neoplasm / Q000378:metabolism* / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D005374:Filtration; D006863:Hydrogen-Ion Concentration; D007850:Lauric Acids / Q000494:pharmacology; D007939:Leukemia L1210 / Q000378:metabolism; D051379:Mice; D009363:Neoplasm Proteins / Q000378:metabolism*; D012521:Sarcosine / Q000031:analogs & derivatives / Q000494:pharmacology; D014965:X-Rays
D004273:DNA, Neoplasm; D007850:Lauric Acids; D009363:Neoplasm Proteins; D002945:Cisplatin; D012521:Sarcosine