Modulation of GST P1-1 activity by polymerization during apoptosis

J Cell Biochem. 2000 Apr;77(4):645-53.

Abstract

Glutathione S-transferases (GSTs, EC 2.5.1.18) belong to a large family of functionally different enzymes that catalyze the S-conjugation of glutathione with a wide variety of electrophilic compounds including carcinogens and anticancer drugs. Drug resistance may result from reduction in apoptosis of neoplastic cells when exposed to antineoplastic drugs. The c-Jun N-terminal Kinase (JNK) belongs to the family of stress kinases and has been shown to be required for the maximal induction of apoptosis by DNA-damaging agents. Recently, an inhibition of JNK activity by GST P1-1, which was reversed by polymerization induced by oxidative stress, has been reported in 3T3-4A mouse fibroblast cell lines. The finding that GST P1-1 might inhibit JNK activity and that it is frequently highly expressed in tumor tissues suggests its possible implication in "apoptosis resistance" during antineoplastic therapy. We investigated the modulation of GST P1-1 during apoptosis in a neoplastic T-cell line (Jurkat) induced by hydrogen peroxide and etoposide. Apoptosis was paralleled by the appearance of a dimeric form of GST P1-1 on western blotting, associated with an increase in the Km(GSH) and a reduction in GST P1-1 specific activity toward 1-chloro-2,4-dinitrobenzene, which reached statistical significance only in H(2)O(2)-treated cells. Our data seem to suggest that H(2)O(2) and etoposide may partly act through a process of partial inactivation of the GST P1-1, possibly involving the "G" site in the process of dimerization, and thus favoring programmed cell death.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis*
  • Blotting, Western
  • DNA Fragmentation
  • DNA, Complementary / metabolism
  • Dimerization
  • Dinitrochlorobenzene / pharmacology
  • Etoposide / pharmacology
  • Glutathione / metabolism
  • Glutathione S-Transferase pi
  • Glutathione Transferase / chemistry
  • Glutathione Transferase / metabolism*
  • Humans
  • Hydrogen Peroxide / pharmacology
  • Indicators and Reagents / pharmacology
  • Isoenzymes / chemistry
  • Isoenzymes / metabolism*
  • Jurkat Cells
  • Models, Biological
  • Nucleic Acid Synthesis Inhibitors / pharmacology
  • Oxidants / pharmacology
  • Restriction Mapping
  • Reverse Transcriptase Polymerase Chain Reaction
  • Time Factors

Substances

  • DNA, Complementary
  • Dinitrochlorobenzene
  • Indicators and Reagents
  • Isoenzymes
  • Nucleic Acid Synthesis Inhibitors
  • Oxidants
  • Etoposide
  • Hydrogen Peroxide
  • GSTP1 protein, human
  • Glutathione S-Transferase pi
  • Glutathione Transferase
  • Glutathione

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