Intracellular kinetics of iron in reticulocytes: evidence for endosome involvement in iron targeting to mitochondria.
In erythroid cells the vast majority of iron (Fe) released from endosomes must cross both the outer and the inner mitochondrial membranes to reach ferrochelatase that inserts Fe into protoporphyrin IX. In the present study, we developed a method whereby a cohort of 59Fe-transferrin (Tf)-laden endosomal vesicles were generated, from which we could evaluate the transfer of 59Fe into mitochondria. Iron chelators, dipyridyl or salicylaldehyde isonicotinoyl hydrazone (SIH), were able to bind the 59Fe when they were present during a 37 degrees C incubation; however, addition of these agents only during lysis at 4 degrees C chelated virtually no 59Fe. Bafilomycin A1 (which prevents endosome acidification) and succinylacetone (an inhibitor of 5-aminolevulinate dehydratase) prevented endosomal 59Fe incorporation into heme. Importantly, both the myosin light chain kinase inhibitor wortmannin and the calmodulin antagonist, N-(6-aminohexyl)-5-chloro-1-naphthalene-sulfonamide (W-7), caused significant inhibition of 59Fe incorporation from 59Fe-Tf-labeled endosomes into heme, suggesting that myosin is required for Tf-vesicle movement. Our results reaffirm the astonishing efficiency of Tf-derived Fe utilization in hemoglobin (Hb)-producing cells and demonstrate that very little of this Fe is present in a chelatable pool. Collectively, these results are congruent with our hypothesis that a transient endosome-mitochondrion interaction mediates iron transfer between these organelles.
Lady Davis Institute for Medical Research, Jewish General Hospital, Montreal, QC, Canada.
An-Sheng
AS
Zhang
Alex D
AD
Sheftel
Prem
P
Ponka
D015082:2,2'-Dipyridyl / Q000494:pharmacology; D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D000447:Aldehydes / Q000494:pharmacology; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport; D011992:Endosomes / Q000187:drug effects / Q000378:metabolism*; D004910:Erythrocyte Membrane / Q000378:metabolism; D006418:Heme / Q000096:biosynthesis / Q000378:metabolism; D006835:Hydrazones / Q000494:pharmacology; D007501:Iron / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007502:Iron Chelating Agents / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D017494:Proton Pumps / Q000378:metabolism; D011522:Protons; D012156:Reticulocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D013449:Sulfonamides / Q000494:pharmacology; D013696:Temperature; D025262:Vacuolar Proton-Translocating ATPases / Q000378:metabolism; D000077191:Wortmannin
D000447:Aldehydes; D000730:Androstadienes; D006835:Hydrazones; D007502:Iron Chelating Agents; D017494:Proton Pumps; D011522:Protons; D013449:Sulfonamides; C413684:salicylaldehyde isonicotinoyl hydrazone; D006418:Heme; D015082:2,2'-Dipyridyl; C017967:W 7; D007501:Iron; D025262:Vacuolar Proton-Translocating ATPases; D000077191:Wortmannin