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Effect of extracellular hemin on hemoglobin and ferritin content of erythroleukemia cells
Author(s) -
Fibach E.,
Konijn A. M.,
Bauminger R. E.,
Ofer S.,
Rachmilewitz E. A.
Publication year - 1987
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041300321
Subject(s) - hemin , ferritin , hemoglobin , transferrin , extracellular , chemistry , biochemistry , intracellular , microbiology and biotechnology , nuclear chemistry , biology , heme , enzyme
Mouse (MEL) and human (K‐562) erythroleukemia cell lines can be induced to undergo erythroid differentiation, including hemoglobin (Hb) synthesis, by extra cellular hemin. In order to study the effect of extracellular hemin on intracellular ferritin and Hb content, we have used Mossabauer spectroscopy to measure the amount of 57 Fe incorporated into ferritin or Hb and a fluorescent enzyme‐linked immunosorbent assay (ELISA) to measure the ferritin protein content. When K‐562 cells were cultured in the presence of a 57 Fe source either as transferrin or citrate, in the absence of a differentiation inducer, all the intracellular 57 Fe was detected in ferritin. When the cells were cultured in the presence of 57 Fe‐hemin, 57 Fe was found in both ferritin and Hb. 57 Fe in ferritin increased rapidly, and after 2 days it reached a plateau at 5 × 10 −14 g/cell. 57 Fe in Hb increased linearly with time and reached the same value after 12 days. Addition of other iron sources such as iron‐saturated transferrin, iron citrate, or iron ammonium citrate caused a much lower increase in ferritin protein content as compared to hemin. When K‐562 cells were induced by 57 Fe‐hemin in the presence of 56 Fe‐transferrin, 57 Fe was found to be incorporated in equal amounts into both ferritin and Hb. However, when the cells were induced by 56 Fe‐hemin in the presence of 57 Fe‐transferrin, 57 Fe was incorporated only into ferritin, but not into Hb, which contained 56 Fe iron. These results indicate that in K‐562 cells, when hemin is present in the culture medium it is preferentially incorporated into Hb, regardless of the availability of other extra‐or intracellular iron sources such as transferrin in ferritin. In MEL cells induced to differentiate by dimethylsulfoxide (DMSO) a different pattern of iron incorporation was observed; 57 Fe from both transferrin and hemin was found to incorporate in ferritin as well as in Hb.