Inactivation of iron responsive element-binding capacity and aconitase function of iron regulatory protein-1 of skin cells by ultraviolet A

Photochem Photobiol. 2000 Dec;72(6):746-52. doi: 10.1562/0031-8655(2000)072<0746:ioireb>2.0.co;2.

Abstract

The ultraviolet-A (UVA) component of sunlight produces in cutaneous cells a highly toxic oxidative stress mediated by redox cycling reactions of Fe ions. A tight regulation of cell iron uptake and storage by iron regulatory proteins (IRP) of keratinocytes and fibroblasts avoids these damaging reactions. We report here that about 40 J/cm2 of UVA are required to inactivate half of the binding capacity of apo-IRP-1 to iron responsive elements (IRE) of RNA whereas 15 J/cm2 already inhibit half of the holo-IRP-1 aconitase activity. No increase in the holo-IRP-1 activity is observed during the apo-IRP-1 photoinactivation suggesting that UVA does not trigger a shift between these two forms. As opposed to holo-IRP-1, which contains a 4Fe-4S cluster, apo-IRP-1 has no UVA chromophore. Thus it should be inactivated indirectly by reactive oxygen species generated by the UVA-induced endogenous photo-oxidative stress. The apo-IRP-1 photoinactivation is weakly prevented by the lipophilic oxyradical scavenger vitamin E but not by the hydrophilic azide anion, a singlet oxygen quencher or by diethyldithiocarbamate, a superoxide dismutase inhibitor. However, full protection against photoinactivation of the apo form is observed after incubation with N-acetylcysteine but the latter only partially protects the aconitase function of the holo-IRP-1 from photoinactivation. The marked difference in the kinetics of photoinactivation of the apo and holo forms, the light dose-independent effect of the sulfhydril group reagent, 2-mercaptoethanol and the partial protection brought by the ferric ion complexing agent desferrioxamine suggest that the photochemistry of the 4Fe-4S cluster of the holo form plays little, if any, role in the photoinactivation of the apo-IRP-1/IRE interaction. It is concluded that the apo/holo equilibrium is irreversibly destroyed by UVA irradiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aconitate Hydratase / metabolism*
  • Cells, Cultured
  • Fibroblasts / enzymology
  • Fibroblasts / metabolism
  • Fibroblasts / radiation effects
  • Humans
  • Iron-Regulatory Proteins
  • Iron-Sulfur Proteins / metabolism*
  • Keratinocytes / enzymology
  • Keratinocytes / metabolism
  • Keratinocytes / radiation effects
  • RNA-Binding Proteins / metabolism*
  • Skin / cytology
  • Skin / enzymology
  • Skin / metabolism
  • Skin / radiation effects*
  • Ultraviolet Rays*

Substances

  • Iron-Regulatory Proteins
  • Iron-Sulfur Proteins
  • RNA-Binding Proteins
  • Aconitate Hydratase