title: Iron dependence of tryptophan hydroxylase activity in RBL2H3 cells and its manipulation by chelators.
abstract: Tryptophan hydroxylase requires Fe2+ for in vitro enzyme activity. In this study, the intracellular activity of tryptophan hydroxylase was assessed by applying 3-hydroxybenzylhydrazine (NSD-1015), an inhibitor of aromatic l-amino acid decarboxylase, to monolayer cultures of RBL2H3 cells, a serotonin producing mast cell line. The effect of manipulating intracellular 'free' iron levels on enzyme activity was analyzed by administration of iron chelators. Desferrioxamine (DFO) suppressed the intracellular enzyme activity. Salicylaldehyde isonicotinoyl hydrazone (SIH) also suppressed enzyme activity, but stimulated it when administered in the Fe-bound form. Hemin also stimulated enzyme activity, which progressively increased over several hours to more than sixfold the initial level. DFO and SIH inhibited the hemin stimulatory effect when administered simultaneously with hemin. Both suppression and stimulation with these chelators took place without a significant decrease or increase in the amount of enzyme. These results indicate that there was an inadequate supply of Fe2+ in the cells to support full activity of tryptophan hydroxylase.
authors.affiliation: Department of Bioscience, Teikyo University of Science and Technology, Yamanashi, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Hasegawa
authors.forename: K
authors.initials: K
authors.lastname: Oguro
authors.forename: Y
authors.initials: Y
authors.lastname: Naito
authors.forename: A
authors.initials: A
authors.lastname: Ichiyama
mesh_terms: D000818:Animals; D002474:Cell-Free System; D003676:Deferoxamine / Q000494:pharmacology; D004789:Enzyme Activation; D006427:Hemin / Q000494:pharmacology; D007501:Iron / Q000378:metabolism*; D007502:Iron Chelating Agents / Q000494:pharmacology*; D051381:Rats; D014365:Tryptophan Hydroxylase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D007502:Iron Chelating Agents; D006427:Hemin; D007501:Iron; D014365:Tryptophan Hydroxylase; D003676:Deferoxamine