Interference by cellular melanin with assay of DNA-protein crosslinks by the potassium dodecyl sulfate precipitation method.
The potassium dodecyl sulfate precipitation method was used to quantify DNA-protein crosslinks (DPCs) in lysates of melanoma cells exposed to ultraviolet radiation. Inducing melanin production in these cells before exposure to ultraviolet radiation decreased the apparent yield of DPCs. The decrease could also be produced by addition of melanin to lysates after exposure to crosslinking conditions. Experimental models could attribute this decrease to neither quenching of scintillations from the tritium label used nor to an effect of single strand breaks of DNA. This assay appears to be inappropriate for quantification of DPCs in melanized cells.
Goodman
R M
RM
Department of Radiology, New Jersey Medical School, Newark 07103-2714, USA.
Hill
H Z
HZ
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003432:Cross-Linking Reagents; D004247:DNA / Q000032:analysis / Q000737:chemistry* / Q000528:radiation effects; D008543:Melanins / Q000494:pharmacology*; D009074:Melanocyte-Stimulating Hormones / Q000494:pharmacology; D008546:Melanoma, Experimental / Q000737:chemistry; D051379:Mice; D011189:Potassium Chloride / Q000494:pharmacology; D011506:Proteins / Q000032:analysis / Q000737:chemistry*; D012967:Sodium Dodecyl Sulfate; D014316:Tritium; D014407:Tumor Cells, Cultured; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D008543:Melanins; D011506:Proteins; D014316:Tritium; D012967:Sodium Dodecyl Sulfate; D011189:Potassium Chloride; D009074:Melanocyte-Stimulating Hormones; D004247:DNA; C028913:dodecyl sulfate; D015056:1-Methyl-3-isobutylxanthine