DNA-binding properties of the antibody specific for the Dewar photoproduct of thymidylyl-(3-5')-thymidine.
A monoclonal antibody (DEM-1) specific for the Dewar photoproduct is used for detection and quantification of photolesions in DNA. To help understand the molecular recognition of damaged DNA by the antibody protein, we have cloned and sequenced the variable region genes of DEM-1. We have also prepared Fab fragments of DEM-1 (DEM1Fab), and synthesized two kinds of 3'-biotinylated oligonucleotides of different lengths containing a central Dewar photoproduct of TpT to analyze the effects of the antigen size on the binding rates by means of surface plasmon resonance (SPR). Results obtained from SPR analyses suggest that DEM1Fab may recognize tetranucleotide unit as the epitope.
Morioka
Hiroshi
H
Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo, Japan. morioka@pahrm.hokudai.ac.jp
Kurihara
Mikiko
M
Kobayashi
Hiroyuki
H
Satou
Kousuke
K
Komatsu
Yasuo
Y
Uchida
Makiyo
M
Ohtsuka
Eiko
E
Torizawa
Takuya
T
Kato
Koichi
K
Shimada
Ichio
I
Matsunaga
Tsukasa
T
Nikaido
Osamu
O
D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D004247:DNA / Q000737:chemistry* / Q000276:immunology* / Q000528:radiation effects; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010777:Photochemistry; D013936:Thymidine / Q000031:analogs & derivatives / Q000737:chemistry / Q000276:immunology* / Q000528:radiation effects*
D000906:Antibodies; D007140:Immunoglobulin Fab Fragments; D004247:DNA; D013936:Thymidine