Immunochemical quantitation of adducts induced in DNA by cis-diamminedichloroplatinum (II) and analysis of adduct-related DNA-unwinding. 1985

A M Fichtinger-Schepman, and R A Baan, and A Luiten-Schuite, and M van Dijk, and P H Lohman

Antibodies elicited against the haptens cis-Pt(NH3)2dGuodGMP and its ribo-analog, both covalently coupled to bovine serum albumin, recognize adducts of cis-diamminedichloroplatinum(II) (cis-DDP) in DNA. Antibody-binding to cis-DDP-DNA strongly depends on the accessibility of the adducts to the antibodies. In double-stranded cis-DDP-DNA with low Pt: nucleotide ratios (rb's), this accessibility is enhanced by unwinding of the cis-DDP-DNA, e.g. by heat-denaturation. An unwinding effect is also induced by the cis-DDP treatment itself. A260nm readings of cis-DDP-DNA samples indicate an increased denaturation of the DNA at increasing Pt-contents. The data obtained after heat-denaturation of the same samples show a growing capability to renaturation when the rb-values increase from 0 to 0.04; at 0.04 less than rb less than 0.18 the renaturation effect gradually disappears. In the competitive enzyme-linked immunosorbent assay (ELISA), the cis-DDP-adducts in heat-denatured DNA are detected in the pmol range; in DNA-digests, however, they are recognized in fmol amounts. For the individual Pt-containing (oligo)nucleotides the amounts causing 50% inhibition in the ELISA were established for the two anti-sera; they were 13.3 +/- 3.8 (fmol +/- S.D.) and 5.4 +/- 1.8 for cis-Pt(NH3)2d(GMP)2; 15.5 +/- 5.4 and 4.0 +/- 1.5 for cis-Pt(NH3)2d(pGpG); (2.6 +/- 1.1) X 10(3) and (2.0 +/- 1.0) X 10(3) for cis-Pt(NH3)2d(pApG); (5.6 +/- 1.9) X 10(3) and (2.9 +/- 0.4) X 10(3) for Pt(NH3)3dGMP. Pt-adducts in a trans-DDP-DNA digest are recognized in pmol amounts and dGMP in nmol quantitatives. Finally, the usefulness of these antibodies for the detection and quantitation of individual cis-DDP-adducts in cis-DDP-DNA digests was demonstrated.

UI MeSH Term Description Entries
D007120 Immunochemistry Field of chemistry that pertains to immunological phenomena and the study of chemical reactions related to antigen stimulation of tissues. It includes physicochemical interactions between antigens and antibodies.
D009691 Nucleic Acid Denaturation Disruption of the secondary structure of nucleic acids by heat, extreme pH or chemical treatment. Double strand DNA is "melted" by dissociation of the non-covalent hydrogen bonds and hydrophobic interactions. Denatured DNA appears to be a single-stranded flexible structure. The effects of denaturation on RNA are similar though less pronounced and largely reversible. DNA Denaturation,DNA Melting,RNA Denaturation,Acid Denaturation, Nucleic,Denaturation, DNA,Denaturation, Nucleic Acid,Denaturation, RNA,Nucleic Acid Denaturations
D011817 Rabbits A burrowing plant-eating mammal with hind limbs that are longer than its fore limbs. It belongs to the family Leporidae of the order Lagomorpha, and in contrast to hares, possesses 22 instead of 24 pairs of chromosomes. Belgian Hare,New Zealand Rabbit,New Zealand Rabbits,New Zealand White Rabbit,Rabbit,Rabbit, Domestic,Chinchilla Rabbits,NZW Rabbits,New Zealand White Rabbits,Oryctolagus cuniculus,Chinchilla Rabbit,Domestic Rabbit,Domestic Rabbits,Hare, Belgian,NZW Rabbit,Rabbit, Chinchilla,Rabbit, NZW,Rabbit, New Zealand,Rabbits, Chinchilla,Rabbits, Domestic,Rabbits, NZW,Rabbits, New Zealand,Zealand Rabbit, New,Zealand Rabbits, New,cuniculus, Oryctolagus
D002853 Chromatography, Liquid Chromatographic techniques in which the mobile phase is a liquid. Liquid Chromatography
D002945 Cisplatin An inorganic and water-soluble platinum complex. After undergoing hydrolysis, it reacts with DNA to produce both intra and interstrand crosslinks. These crosslinks appear to impair replication and transcription of DNA. The cytotoxicity of cisplatin correlates with cellular arrest in the G2 phase of the cell cycle. Platinum Diamminodichloride,cis-Diamminedichloroplatinum(II),cis-Dichlorodiammineplatinum(II),Biocisplatinum,Dichlorodiammineplatinum,NSC-119875,Platidiam,Platino,Platinol,cis-Diamminedichloroplatinum,cis-Platinum,Diamminodichloride, Platinum,cis Diamminedichloroplatinum,cis Platinum
D004247 DNA A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine). DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D006241 Haptens Small antigenic determinants capable of eliciting an immune response only when coupled to a carrier. Haptens bind to antibodies but by themselves cannot elicit an antibody response. Hapten,Contact-Sensitizing Agents,Agents, Contact-Sensitizing,Contact Sensitizing Agents
D006358 Hot Temperature Presence of warmth or heat or a temperature notably higher than an accustomed norm. Heat,Hot Temperatures,Temperature, Hot,Temperatures, Hot
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia

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