Activation of in situ tissue transglutaminase by intracellular reactive oxygen species

Biochem Biophys Res Commun. 2003 Jun 6;305(3):633-40. doi: 10.1016/s0006-291x(03)00835-0.

Abstract

We have investigated the novel function of intracellular reactive oxygen species (ROS) in the activation of in situ tissue transglutaminase (tTGase) by lysophosphatidic acid (LPA) and transforming growth factor-beta (TGF-beta) in Swiss 3T3 fibroblasts. LPA induced a transient increase of intracellular ROS with a maximal increase at 10 min, which was blocked by ROS scavengers, N-acetyl-L-cysteine and catalase. LPA activated tTGase with a maximal increase at 1h, which was inhibited by cystamine and ROS scavengers. Incubation with exogenous H(2)O(2) activated tTGase. TGF-beta also activated tTGase with a maximal activation at 2h and the tTGase activation was inhibited by the ROS scavengers. Scrape-loading of C3 transferase inhibited the ROS production and in situ tTGase activation by LPA and TGF-beta, and the inhibitory effect of C3 transferase was reversed by exogenous H(2)O(2). Microinjection of GTPgammaS inhibited transamidating activity of tTGase stimulated by LPA, TGF-beta, and maitotoxin. These results suggested that intracellular ROS was essential for the activation of in situ tTGase in response to LPA and TGF-beta.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP Ribose Transferases / pharmacology
  • Animals
  • Botulinum Toxins / pharmacology
  • Cell Line
  • Enzyme Activation
  • Enzyme Inhibitors / pharmacology
  • Fibroblasts / drug effects
  • Fibroblasts / enzymology
  • GTP-Binding Proteins / metabolism*
  • Guanosine 5'-O-(3-Thiotriphosphate) / administration & dosage
  • Guanosine 5'-O-(3-Thiotriphosphate) / pharmacology
  • Kinetics
  • Lysophospholipids / antagonists & inhibitors
  • Lysophospholipids / pharmacology*
  • Mice
  • Microinjections
  • Protein Glutamine gamma Glutamyltransferase 2
  • Reactive Oxygen Species / metabolism*
  • Transforming Growth Factor beta / antagonists & inhibitors
  • Transforming Growth Factor beta / pharmacology*
  • Transglutaminases / metabolism*

Substances

  • Enzyme Inhibitors
  • Lysophospholipids
  • Reactive Oxygen Species
  • Transforming Growth Factor beta
  • Guanosine 5'-O-(3-Thiotriphosphate)
  • Protein Glutamine gamma Glutamyltransferase 2
  • Transglutaminases
  • ADP Ribose Transferases
  • exoenzyme C3, Clostridium botulinum
  • Botulinum Toxins
  • GTP-Binding Proteins