Photoaffinity labeling of tumor promoter-binding protein (CN-TPBP) and preparation of affinity sorbent gels.
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.
Azuma
A
A
Institute of Molecular & Cellular Biosciences, University of Tokyo, Bunkyo-ku, Japan.
Ando
Y
Y
Iwasaki
S
S
Hashimoto
Y
Y
Yamaguchi
M
M
Takehana
S
S
Fukasawa
H
H
Endo
Y
Y
Shudo
K
K
D000345:Affinity Labels; D002352:Carrier Proteins / Q000032:analysis*; D005782:Gels; D008970:Molecular Weight; D013755:Tetradecanoylphorbol Acetate / Q000378:metabolism
D000345:Affinity Labels; D002352:Carrier Proteins; D005782:Gels; C082634:cytosolic-nuclear tumor promoter binding protein; D013755:Tetradecanoylphorbol Acetate