title: Apoptotic cell death analyzed at the molecular level by two-dimensional gel electrophoresis.
abstract: The pattern of protein expression and phosphorylation after an apoptotic stimulus has been studied in two systems. Bovine aortic endothelial cells were induced to undergo apoptotic cell death by a combination of a cytokine (tumor necrosis factor, TNF) and inhibitors of protein synthesis, like cycloheximide. Two-dimensional (2-DE) electrophoresis of proteins from such cells revealed specific proteolysis of distinct proteins, some at an early stage of apoptosis and some at a later stage. These proteins may have antiapoptotic properties. In rat IPC-81 promyelocytic leukemia cells, cAMP induced apoptosis. 2-DE of such cells pulse-labeled with [35S]methionine revealed two "novel" protein spots (of 30 kDa and 46 kDa, respectively), induced very rapidly by a posttranscriptional mechanism. It is proposed that "dysphosphorylation" may accompany apoptosis in general, since both endothelial cells treated with TNF/cycloheximide and IPC-81 cells treated with cAMP analog or the apoptosis-inducing phosphatase inhibitors okadaic acid or calyculin A all showed altered protein phosphorylation patterns, as revealed by 2-DE electrophoresis of proteins from cells prelabeled with 32Pi.
authors.affiliation: Institute of Interdisciplinary Research, School of Medicine, Free University of Brussels, Belgium.
authors.forename: B
authors.initials: B
authors.lastname: Robaye
authors.forename: A P
authors.initials: AP
authors.lastname: Døskeland
authors.forename: N
authors.initials: N
authors.lastname: Suarez-Huerta
authors.forename: S O
authors.initials: SO
authors.lastname: Døskeland
authors.forename: J E
authors.initials: JE
authors.lastname: Dumont
mesh_terms: D000818:Animals; D001011:Aorta; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D001345:Autoradiography / Q000379:methods; D002417:Cattle; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D010450:Endopeptidases / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D005453:Fluorescence; D007930:Leucine / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D014176:Protein Biosynthesis; D011506:Proteins / Q000302:isolation & purification; D013873:Thionucleotides / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D010710:Phosphates; D010750:Phosphoproteins; D010761:Phosphorus Radioisotopes; D011506:Proteins; D013873:Thionucleotides; D014409:Tumor Necrosis Factor-alpha; C015929:8-((4-chlorophenyl)thio)cyclic-3',5'-AMP; D000242:Cyclic AMP; D010450:Endopeptidases; D007930:Leucine