Lysosomal and cytosolic ferritins. A biochemical and electron-spectroscopic study

Cytosolic and lysosomal ferritin and haemosiderin were isolated from rat livers which had been iron-loaded by four intraperitoneal injections of iron-dextran. The cytosolic and lysosomal ferritins, prepared in a phosphate-free medium, were subjected to gel-filtration chromatography on Sepharose 6B,...

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Bibliographic Details
Published in:Biology of metals 1989, Vol.2 (2), p.114-121
Main Authors: Ringeling, P L, Cleton, M I, Kroos, M J, Sorber, L W, de Bruyn, W C, Harrison, P M, van Eijk, H G
Format: Article
Language:English
Subjects:
Animals
Chromatography, Gel
Cytosol - analysis
Electron Probe Microanalysis
Ferritins - isolation & purification
Hemosiderin - isolation & purification
Iron - analysis
Liver - analysis
Lysosomes - analysis
Male
Microscopy, Electron
Phosphorus - analysis
Rats
Rats, Inbred Strains
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Summary:Cytosolic and lysosomal ferritin and haemosiderin were isolated from rat livers which had been iron-loaded by four intraperitoneal injections of iron-dextran. The cytosolic and lysosomal ferritins, prepared in a phosphate-free medium, were subjected to gel-filtration chromatography on Sepharose 6B, yielding four fractions: a cytosolic monomeric (CMF) and void-volume ferritin fraction (CVVF), and a lysosomal monomeric (LMF) and void-volume ferritin fraction (LVVF). Of each fraction the following aspects were examined: (a) immunoreactivity against specific antiserum; (b) the Fe/P mass ratio and the effect of dialysis on this ratio using electron probe micro-analysis (EPMA); (c) morphology and Fe-specific imaging using electron spectroscopic imaging (ESI) and electron energy loss spectroscopy (EELS). For haemosiderin one aspect, the Fe/P ratio, was determined before and after extensive purification. The following results were obtained (a) All ferritin fractions reacted with anti- (rat liver ferritin). (b) The Fe/P ratios as determined in CMF in an haemosiderin were not affected by dialysis or extensive purification, respectively. The Fe/P ratio in CVVF was affected by dialysis. In the lysosomal fractions, only a trace of phosphorus (LVVF) or no phosphorus (LMF) was detected. (c) Morphologically, CMF and CVVF were found to be rather homogeneous; the iron core diameters of both fractions were in the known size range.
ISSN:0933-5854
1572-8773
DOI:10.1007/BF01129210