Alterations in cellular Ca(2+) and free iron pool by sulfur amino acid deprivation: the role of ferritin light chain down-regulation in prooxidant production.
Deficiency of sulfur amino acids occurs in certain pathophysiological states such as protein-calorie malnutrition. Sulfur amino acid deprivation (SAAD) increases oxidative stress through a decrease in GSH. Ferritin expression is induced by oxidative stress, which confers resistance to oxidative insults. The effects of SAAD on the changes in cellular Ca(2+) and free iron pool, prooxidant production and the ferritin light chain (FLC) expression were comparatively evaluated in Hepa1c1c7 and Raw264.7 cells. [Ca(2+)](i) was rapidly increased by SAAD. Sulfhydryl-containing compounds prevented the increase in [Ca(2+)](i) in cells under SAAD, supporting the role of redox-state in the regulation of [Ca(2+)](i). Thapsigargin or Ca(2+)-free medium inhibited the increase in [Ca(2+)](i), showing that Ca(2+) originated from endoplasmic reticulum as well as from extracellular source. Inhibition of Ca(2+) mobilization decreased the fluorescence of Phen Green SK inside cells, representing the inhibition of free iron release. Both inhibition of Ca(2+) mobilization and iron chelation decreased dichlorofluorescein oxidation, indicating the possibility that the increase in [Ca(2+)](i) affected that in cellular free iron and prooxidant production. FLC protein level was immunochemically detectable in Raw264.7 cells, but not in Hepa1c1c7 cells. SAAD alone (or in combination with FeSO(4)) down-regulated FLC protein expression, while SAAD increased the FLC mRNA level in both Hepa1c1c7 and Raw264.7 cells. Calcium or iron chelators prevented increases in the FLC mRNA. These results provided evidence that changes in cellular Ca(2+) and iron pool by SAAD increased cellular oxidative stress and that the down-regulation of FLC protein by SAAD would further enhance prooxidant production in spite of the increase in FLC mRNA.
Kim
Hye Jung
HJ
College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, 151-742, Seoul, South Korea.
Kim
Sang Geon
SG
D000603:Amino Acids, Sulfur / Q000172:deficiency / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D015536:Down-Regulation; D005293:Ferritins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D007501:Iron / Q000378:metabolism*; D051379:Mice; D016877:Oxidants / Q000378:metabolism*; D018384:Oxidative Stress; D012333:RNA, Messenger / Q000096:biosynthesis
D000603:Amino Acids, Sulfur; D016877:Oxidants; D012333:RNA, Messenger; D005293:Ferritins; D007501:Iron; D002118:Calcium