Premium
Apoptotic cell death analyzed at the molecular level by two‐dimensional gel electrophoresis
Author(s) -
Robaye Bernard,
Døskeland Anne Paulus,
SuarezHuerta Nathalie,
Døskeland Stein Ove,
Dumont Jacques E.
Publication year - 1994
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150150168
Subject(s) - cycloheximide , apoptosis , biology , programmed cell death , microbiology and biotechnology , phosphorylation , biochemistry , protein biosynthesis
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 [ 35 S]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 32 Pi.