Azuma A, Ando Y, Iwasaki S, Hashimoto Y, Yamaguchi M, Takehana S, Fukasawa H, Endo Y, Shudo K
Institute of Molecular & Cellular Biosciences, University of Tokyo, Bunkyo-ku, Japan.
Biol Pharm Bull. 1997 Jan;20(1):6-9. doi: 10.1248/bpb.20.6.
Cytosolic-nuclear tumor promoter-specific binding protein (CN-TPBP) was photoaffinity-labeled with a specific ligand, 3beta,5alpha-dihydroxycholestan-6-one (YS-64). Analysis by ODS-HPLC of peptide fragments obtained from the labeled CN-TPBP by trypsinization indicated the existence of a single specifically labeled site. Affinity gels for the purification of CN-TPBP were then prepared. As ligands for the affinity gels, 12-O-tetradecanoylphorbol 13-acetate (TPA), a typical phorbol-type tumor promoter, and benzolactam-V8-310 (BL-V8-310), a structural/biological mimic of teleocidin-class tumor promoters, were adopted. The use of these gels afforded a protein that showed a single band of 58 kDa on SDS-PAGE.
