Antigen structural requirements for recognition by a cyclobutane thymine dimer-specific monoclonal antibody.
A monoclonal antibody (TDM-2) specific to a UV-induced cyclobutane pyrimidine dimer (T[cis-syn]T) has previously been established; however,the immunization had used UV-irradiated calf-thymus DNA containing a heterogeneous mixture of photoproduct sites. We investigated here the structural requirements of antigen recognition by the antibody using chemically synthesized antigen analogs. TDM-2 bound with cis-syn,but not trans-syn thymine dimer,and could bind strongly with four nucleotide analogs in which the cis-syn pyrimidine dimer was located in the center. Antigen analogs containing abasic linkers at the 5'- or 3'-side of the cis-syn cyclobutane pyrimidine dimer were synthesized and tested for binding to TDM-2. The results indicated that TDM-2 recognizes not only the cyclobutane ring but also both the 5'- and 3'-side nucleosides of the cyclobutane dimer. Furthermore,it was proved that either the 5'- or 3'-side phosphate group at a cyclobutane dimer site was absolutely required for the affinity to TDM-2. The antibody showed a strong binding to single stranded DNA but indicated little binding to double stranded DNA.
Komatsu
Y
Y
Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo 060, Japan.
Tsujino
T
T
Suzuki
T
T
Nikaido
O
O
Ohtsuka
E
E
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000941:Antigens / Q000737:chemistry*; D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D002417:Cattle; D004247:DNA / Q000737:chemistry / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D066298:In Vitro Techniques; D051379:Mice; D015394:Molecular Structure; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D011740:Pyrimidine Dimers / Q000138:chemical synthesis / Q000737:chemistry* / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000941:Antigens; D009838:Oligodeoxyribonucleotides; D011740:Pyrimidine Dimers; D004247:DNA