Release of intact, monomeric cytochrome c from apoptotic and necrotic cells.
Cytochrome c (Cyt c) has been shown to translocate from mitochondria to the cytoplasm of cells early in apoptosis. In this study sandwich ELISAs for Cyt c were used to determine if Cyt c is ultimately released from apoptotic and necrotic cells. Gel-filtration and cation-exchange chromatographies, in conjunction with immunoreactivity in ELISA, and Western blotting were employed to examine the integrity of the released Cyt c. The results show that Cyt c is released from both apoptotic and necrotic cells in an intact, monomeric form. The release of Cyt c from apoptotic splenocytes began within 2 h following apoptotic insult, while Cyt c was immediately released following induction of necrosis by heat shock. These findings may be relevant to understanding how Cyt c becomes a target for antibody production in some patients with systemic autoimmune diseases.
Jemmerson
R
R
Department of Microbiology and Center for Immunology, University of Minnesota Medical School, Minneapolis, MN 55455, USA. ron@lenti.med.umn.edu
LaPlante
B
B
Treeful
A
A
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D020126:Brefeldin A / Q000494:pharmacology; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D003574:Cytochrome c Group / Q000378:metabolism*; D004789:Enzyme Activation; D018922:HL-60 Cells / Q000187:drug effects; D018882:Heat Stress Disorders / Q000473:pathology; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D051379:Mice; D009336:Necrosis; D013154:Spleen / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology / Q000502:physiology; D014407:Tumor Cells, Cultured
D016895:Culture Media, Serum-Free; D003574:Cytochrome c Group; D020126:Brefeldin A; D007770:L-Lactate Dehydrogenase; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases