Release of intact, monomeric cytochrome c from apoptotic and necrotic cells

Cell Death Differ. 2002 May;9(5):538-48. doi: 10.1038/sj.cdd.4400981.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Apoptosis / physiology*
  • Brefeldin A / pharmacology
  • Caspase 3
  • Caspases / metabolism
  • Culture Media, Serum-Free / pharmacology
  • Cytochrome c Group / metabolism*
  • Enzyme Activation
  • HL-60 Cells / drug effects
  • Heat Stress Disorders / pathology
  • Humans
  • L-Lactate Dehydrogenase / metabolism
  • Mice
  • Necrosis
  • Spleen / drug effects
  • Spleen / enzymology*
  • Spleen / pathology
  • Spleen / physiology
  • Tumor Cells, Cultured

Substances

  • Culture Media, Serum-Free
  • Cytochrome c Group
  • Brefeldin A
  • L-Lactate Dehydrogenase
  • CASP3 protein, human
  • Casp3 protein, mouse
  • Caspase 3
  • Caspases