In situ photoaffinity labeling of proteasome with photoactive adriamycin analogue

Biochem Biophys Res Commun. 2000 Jul 14;273(3):928-32. doi: 10.1006/bbrc.2000.3027.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cysteine Endopeptidases / chemistry*
  • Cysteine Endopeptidases / isolation & purification
  • Doxorubicin / analogs & derivatives*
  • Doxorubicin / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Leukemia L1210 / pathology
  • Mice
  • Multienzyme Complexes / chemistry*
  • Multienzyme Complexes / isolation & purification
  • Photoaffinity Labels
  • Proteasome Endopeptidase Complex
  • Tumor Cells, Cultured

Substances

  • Multienzyme Complexes
  • Photoaffinity Labels
  • Doxorubicin
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex