In situ photoaffinity labeling of proteasome with photoactive adriamycin analogue.
An intracellular adriamycin (ADM)-binding protein purified from the cytosol of L1210 mouse lymphocytic leukemia cells had a molecular weight of 700-1500 kDa and hydrolyzed Suc-LLVY-MCA. When L1210 cells were incubated with a photoactive ADM analogue, N-(p-azidobenzoyl)-adriamycin (NAB-ADM), most of the NAB-ADM was found to localize in the nuclei. In situ photoaffinity labeling of L1210 cells with NAB-ADM resulted in low protease activity in the cytosol and nuclear extracts and the cells showed selective photoincorporation of NAB-ADM into the proteasome. These results suggest that the proteasome is a translocator of ADM from the cytoplasm to the nucleus and might therefore become a new candidate for cancer chemotherapy.
Kiyomiya
K I
KI
Department of Toxicology, Osaka Prefecture University, Sakai, Osaka, 599-8531, Japan. kiyomiya@vet.osakafu-u.ac.jp
Matsuo
S
S
Kurebe
M
M
D000818:Animals; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification; D004317:Doxorubicin / Q000031:analogs & derivatives* / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D007939:Leukemia L1210 / Q000473:pathology; D051379:Mice; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000302:isolation & purification; D019920:Photoaffinity Labels; D046988:Proteasome Endopeptidase Complex; D014407:Tumor Cells, Cultured
D009097:Multienzyme Complexes; D019920:Photoaffinity Labels; D004317:Doxorubicin; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex