Maud I. Cleton

Structure and composition of ferritin cores from pea seed (Pisum sativum)

Iron cores from native pea seed (Pisum sativum) ferritin have been analysed by electron microscopy and Mössbauer spectroscopy and shown to be amorphous. This correlates with their relatively high phosphate content (Fe: P = 2.83; 1800 Fe, 640 P atoms/molecule). Reconstituted cores obtained by adding iron (2000 Fe atoms/molecule) in the absence of phosphate to pea seed apoferritin were crystalline ferrihydrite. In vitro rates of formation of pea-seed ferritin iron cores were intermediate between those of recombinant human H-chain and horse spleen apoferritin and this may reflect the amino-acid residues of its ferroxidase and putative nucleation centres. The high phosphate content of pea-seed ferritin suggests that this molecule could be involved in both phosphorus and iron storage. The high phosphate concentration found within plastids, from which the molecules were isolated, is a possible source of the ferritin phosphate.

A note on the composition and properties of ferritin iron cores

In ferritins and bacterioferritins iron is stored as an inorganic complex within a protein shell. The composition and properties of this complex are surprisingly variable. Factors that may lead to such variability are discussed.

Studies on haemosiderin and ferritin from iron-loaded rat liver

SummaryHaemosiderin has been isolated from siderosomes and ferritin from the cytosol of livers of rats iron-loaded by intraperitoneal injections of iron-dextran. Siderosomal haermosiderin, like ferritin, was shown by electron diffraction to contain iron mainly in the form of small particles of ferrihydrite (5Fe2O3 · 9H2O), with average particle diameter of 5.36±1.31 nm (SD), less than that of ferritin iron-cores (6.14±1.18 nm). Mössbauer spectra of both iron-storage complexes are also similar, except that the blocking temperature,TB, for haemosiderin (23 K) is lower than that of ferritin (35 K). These values are consistent with their differences in particle volumes assuming identical magnetic anisotropy constants. Measurements of P/Fe ratios by electron probe microanalysis showed the presence of phosphorus in rat liver haemosiderin, but much of it was lost on extensive dialysis. The presence of peptides reacting with anti-ferritin antisera and the similarities in the structures of their iron components are consistent with the view that rat liver haemosiderin arises by degradation of ferritin polypeptides, but its peptide pattern is different from that found in humanβ-thalassaemia haemosiderin. The blocking temperature, 35 K, for rat liver ferritin is near to that reported, 40 K, for humanβ-thalassaemia spleen ferritin. However, the haemosiderin isolated from this tissue, in contrast to that from rat liver, had aTB higher than that of ferritin. The iron availability of haemosiderins from rat liver and humanβ-thalassaemic spleen to a hydroxypyridinone chelator also differed. That from rat liver was equal to or greater, and that from human spleen was markedly less, than the iron availability from either of the associated ferritins, which were equivalent. The differences in properties of the two types of haemosiderin may reflect their origins from primary or secondary iron overload and differences in the duration of the overload.

Studies on ferritin in rat liver and spleen during repeated phlebotomy

1. The ferritin content of liver and spleen in normal and iron-loaded rats decreased during repeated phlebotomy. 2. During increased iron demand, ferritin is degraded in toto. 3. With the ESI and EELS technique the iron distribution was followed in different cell types and cellular compartments. 4. We have demonstrated two methods of iron mobilisation: (a) catabolism of lysosomal ferritin in toto and (b) delivery of ferritin from parenchymal cell into the bile and degradation of ferritin in toto.

Effect of phlebotomy on the ferritin iron content in the rat liver as determined morphometrically with the use of electron energy loss spectroscopy

SummaryPhlebotomy of untreated and iron-loaded rats results in a significant decrease in total liver iron. In ironloaded rats a marked decrease in iron-containing particles is observed ultrastructurally in lysosomes and cytoplasm of hepatic sinusoidal cells but not in parenchymal cells. This remarkable phenomenon was further investigated in a morphometric study, based on element-specific (iron) distribution images made in situ in the parenchymal cell by means of electron energy loss spectroscopy. With the use of this technique it could be shown that in spite of phlebotomy the ferritin iron content of the iron-loaded liver parenchymal cell is not decreased.

Lysosomal and cytosolic ferritins A biochemical and electron-spectroscopic study

SummaryCytosolic 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 613, 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 Fespecific 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 CWF 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. LMF and LVVF were of rather heterogeneous composition; the core diameters of these fractions were different. In conclusion: the phosphorus in ferritin and haemosiderin is firmly bound; Haemosiderin, when derived from ferritin, has to take up phosphorus in the lysosomes.

Comparison of cytosolic products formed in rat liver in response to parenteral and dietary iron loading.

Two different methods were used to create a situation of iron (Fe) overload in rats. One group of rats received Fe dextran, and another group of rats received a carbonyl Fe-enriched diet. The ferritins present in the liver cytosol of these rats were isolated and compared. From each group, two cytosolic products were isolated with the use of ultracentrifugation: a cytosolic ferritin fraction (CF) and a (slower sedimenting) light ferritin fraction (CLF). There were no differences with respect to the protein coat (subunit composition and amino acid analysis). Analysis of the Fe core revealed that the two CF fractions were similar, whereas the two CLF fractions differed with respect to their Fe content and to the packing of their cores. The carbonyl CLF product contained less Fe atoms/molecule, which, moreover, seemed to be packed in a less compact way.

Clinical chemistry in the medical curriculum: From practical to research

The haemoglobin levels of a group of healthy students aged 18-3(1 years (n = 216) were determined. These were matched with a test population of 34 wind instrument players. The average haemoglobin level of the wind players was significantly higher.