Antibodies for denatured human H-ferritin stain only reticuloendothelial cells within the bone marrow.
Human H-ferritin homopolymer was denatured in sodium dodecyl sulphate and injected in mice to obtain antibodies for dissociated H-subunit. The antisera and Moabs obtained were specific for the denatured H-chain with no cross-reactivity with assembled ferritins in immunoblotting experiments. In contrast the Moabs for native recombinant H-ferritin are specific for the assembled ferritin molecules with weak cross-reactivity with the denatured H-subunits. The epitope recognized by one of the anti-denatured H-chain Moabs was mapped on the C-terminal helix of ferritin. The antibodies were used to study H-ferritin conformation in cells. In immunocytochemistry experiments the antibodies for denatured H-ferritin stained HeLa and K562 cells weakly, with a different intensity and pattern to those obtained with anti-native H-ferritin antibody. In human bone marrow smears the anti-denatured ferritin antibodies stained only reticuloendothelial cells, and did not recognize the H-ferritin rich immature erythroblasts. It is concluded that assembled and denatured H-ferritins are immunogenically distinct, and that erythroid and reticuloendothelial cells within the bone marrow contain H-ferritin in different conformations.
Ruggeri
G
G
Institute of Chemistry, University of Brescia, Italy.
Santambrogio
P
P
Bonfiglio
F
F
Levi
S
S
Bugari
G
G
Verardi
R
R
Cazzola
M
M
Invernizzi
R
R
Zambelli
L M
LM
Albertini
A
A
D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D001853:Bone Marrow / Q000737:chemistry / Q000276:immunology; D001854:Bone Marrow Cells; D004591:Electrophoresis, Polyacrylamide Gel; D015672:Erythroid Precursor Cells / Q000737:chemistry / Q000166:cytology / Q000276:immunology; D005293:Ferritins / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D007501:Iron / Q000378:metabolism; D012157:Mononuclear Phagocyte System / Q000737:chemistry* / Q000166:cytology*; D011489:Protein Denaturation; D013194:Staining and Labeling
D000911:Antibodies, Monoclonal; D005293:Ferritins; D007501:Iron