Mutated recombinant human heavy-chain ferritins and myelosuppression in vitro and in vivo: a link between ferritin ferroxidase activity and biological function.
Human heavy-chain (H-) ferritin muteins obtained by oligonucleotide site-directed mutagenesis, together with wild-type recombinant human H- and light-chain (L-) ferritins, were evaluated for in vitro effects on the suppression of human bone marrow myeloid progenitor cells and for in vivo effects on marrow and splenic myelopoiesis in C3H/HeJ mice. The 10 H-ferritin muteins exhibited alterations of various regions of the molecule, including ones exposed on the outer surface, on the inner cavity, and on the hydrophilic and hydrophobic channels and of the four-alpha-helix bundle forming the subunit structure. They were stable and were electrophoretically analogous to wild-type H-ferritin. The muteins showed in vitro and in vivo myelosuppressive activity analogous to wild type, except for mutein 222, which was totally inactive and which lacked ferroxidase activity. Recombinant human L-ferritin, devoid of ferroxidase activity, was also inactive as a suppressor. The results demonstrate that H-ferritin myelosuppressive and ferroxidase activities are linked. One possibility is that ferroxidase activity may interfere with the cellular uptake of transferrin iron that is needed for cell proliferation, an interpretation consistent with the presently described ability of hemin to overcome H-ferritin suppressive effects.
Broxmeyer
H E
HE
Department of Medicine, Hematology/Oncology, Indiana University School of Medicine, Indianapolis 46202.
Cooper
S
S
Levi
S
S
Arosio
P
P
D000818:Animals; D001853:Bone Marrow / Q000502:physiology*; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D005293:Ferritins / Q000235:genetics / Q000494:pharmacology*; D006410:Hematopoiesis / Q000187:drug effects; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D051379:Mice; D008809:Mice, Inbred C3H; D016297:Mutagenesis, Site-Directed; D010088:Oxidoreductases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology; D013329:Structure-Activity Relationship
D011994:Recombinant Proteins; D005293:Ferritins; D010088:Oxidoreductases; C023100:ferroxidase II