Binding of cis- and trans-diamminedichloroplatinum(II) to deoxyribonucleic acid exposes nucleosides as measured immunochemically with anti-nucleoside antibodies.
We report the use of anti-nucleoside antibodies to probe for local denaturation of calf thymus DNA upon binding of the antitumor drug cis-diamminedichloroplatinum(II), cis-DDP, and the biologically inactive analogues trans-diamminedichloroplatinum(II), trans-DDP, and chloro(diethylenetriamine)platinum(II) chloride, [Pt(dien)Cl]Cl. These antibodies specifically recognize each of the four DNA nucleosides. They bind well to denatured DNA, but not to native DNA in which the bases are less accessible owing to Watson-Crick duplex structure. At relatively high levels of modification (D/N approximately 0.1), cis-DDP causes significant disruption of DNA base pairing as reflected by the increased binding of anti-cytidine, anti-adenosine, and anti-thymidine antibodies. At lower levels of platinum adduct formation, however, all four anti-nucleoside antibodies bind more to DNA modified with trans-DDP. This result indicates that adducts formed by trans-DDP disrupt the DNA structure to a greater extent than those formed by cis-DDP at low D/N ratios. Modification of DNA by the monofunctional complex [Pt(dien)Cl]Cl does not affect its recognition by anti-nucleoside antibodies, demonstrating that base pair disruption is a consequence of bifunctional binding. The relative anti-nucleoside antibody recognition of cis-DDP-modified DNA is anti-cytosine greater than anti-adenosine approximately anti-thymidine much greater than anti-guanosine, consistent with the major adduct being an intrastrand d(GpG) cross-link. These results reveal that base pair disruption in a naturally occurring DNA modified by either cis-DDP or trans-DDP is sufficient to be detected by protein (antibody) binding. The relevance of these findings to current ideas about the molecular mechanism of action of cis-DDP is discussed.
Sundquist
W I
WI
Lippard
S J
SJ
Stollar
B D
BD
D000818:Animals; D000936:Antigen-Antibody Complex; D002417:Cattle; D002945:Cisplatin / Q000031:analogs & derivatives; D004247:DNA; D003853:Deoxyribonucleosides / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006358:Hot Temperature; D007106:Immune Sera; D007536:Isomerism; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D013329:Structure-Activity Relationship; D013950:Thymus Gland
D000936:Antigen-Antibody Complex; D003853:Deoxyribonucleosides; D007106:Immune Sera; C032799:chlorodiethylenetriamine platinum; C034697:transplatin; D004247:DNA; D002945:Cisplatin