Electrophoresis of Ferritins

Zamiri, Iraj; Mason, Judy

doi:10.1038/217258a0

Cited by 13 publications

(5 citation statements)

References 4 publications

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“…The heterogeneity with respect to the latter state of aggregation is reflected by the appearance of at least three bands of different electrophoretic mobilities on starch or polyacrylamide gels, and the chromatographic resolution of purified ferritin into several fractions (Richter, 1963; Fine and Harris, 1963;Kopp et al, 1963Kopp et al, , 1964Theron et al, 1963; Carnevali and Tecce, 1964). Although multiaggregates have often been observed in various species (Zamiri and Mason, 1968) and in normal and tumor ferritins (Richter and Lee, 1970), the structural basis for this phenomenon was not elucidated. Apoferritin prepared by reduction and removal of the iron from native ferritin is also a heterogeneous mixture containing a main component with a sedimentation coefficient of 17 S and minor components of about 24 and 33 S (Suran and Tarver, 1965;Harrison and Gregory, 1965;Williams et al, 1966).…”

mentioning

confidence: 99%

Mechanisms for the formation of ferritin oligomers

Niitsu¹,

Listowsky²

1973

Biochemistry

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confidence: 99%

Mechanisms for the formation of ferritin oligomers

Niitsu¹,

Listowsky²

1973

Biochemistry

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“…Electrophoresis was carried out in the same buffer for 8 h at a field strength of 3.5 mA/cm. Gels were then fixed overnight in a mixture of 45% methanol and 45% acetic acid, subsequently stained for I h with 0.03% Coomassie Brilliant Blue R-250 in 24% (vol/vol) isopropanol, and 10% (vol/vol) acetic acid, followed by a final destaining wash in the same isopropanol-acetic acid solvent (40,50) (Fig. 2) .…”

mentioning

confidence: 99%

Histamine receptors of the microvascular endothelium revealed in situ with a histamine-ferritin conjugate: characteristic high-affinity binding sites in venules.

Heltianu¹,

Simionescu²,

Simiónescu³

1982

The Journal of Cell Biology

135

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Histamine covalently bound to glutaraldehyde-activated ferritin was prepared as either monomers or as small aggregates of -0 .05 to 0.15 p,m Diam, suitable for electron microscopic detection of histamine cellular binding sites. The histamine-ferritin conjugates (MF) maintain the histamine capability to induce the opening of endothelial junctions in venules. To investigate the distribution of histamine receptors in the vascular endothelium, monomers or aggregates of MF were perfused in situ (mice), and various vascular beds, particularly that of the diaphragm, were fixed and processed for electron microscopy . The conjugate was preferentially bound on restricted areas of luminal endothelial cell plasmalemma especially in regions rich in filaments, and near the junctions between endothelial cells. The density of histamine binding sites was characteristically high in venules; it occurred to a much lesser extent in arterioles, veins, and muscular arteries whereas capillaries and aorta showed the lowest values . A similar distribution was obtained after perfusion of H, or H 2 receptor agonists coupled to ferritin (2-pyridylethylamine-ferritin [PF], or 4-methylhistamine-ferritin [MF], respectively) . The binding specificity was assessed through control experiments with either native or activated ferritin or by competition with histamine. The findings suggest that histamine receptors are largely represented in the cell membrane of the vascular endothelium, particularly in venules. Experiments using specific H, and H2 receptor agonists (PF and MF) and antagonists (mepyramine and cimetidine) indicate that the venular endothelium contains mainly H 2 receptors.

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“…Our results indicate that chicken liver ferritin contains a major componen-t the 470 kDa monomer on nondenaturing PAGE, and a minor dimer component similar to chicken spleen ferritin (Shinjyo, 1973) or bovine ferritin (Zampiri & Mason, 1968). Commercially available horse spleen ferritin was found to consist of a faster-migrating form of 470 kDa (Stefanini et al, 1982).…”

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confidence: 63%

Purification of chicken liver ferritin by two novel methods and structural comparison with horse spleen ferritin

Passaniti

Roth

1989

Biochemical Journal

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Ferritin was purified from chicken liver by two different methods: gel filtration on controlled-pore glass beads, and immunoaffinity chromatography employing a chicken ferritin-specific monoclonal antibody that did not cross-react with horse spleen ferritin. This antibody recognizes intact ferritin and an oligomeric 240 kDa form of the molecule after protein transfer to nitrocellulose, but not the 22 kDa chicken ferritin subunit. Chicken liver ferritin purified by these methods exhibited reduced migration on non-denaturing polyacrylamide gels compared with horse spleen ferritin. These results were consistent with the difference in calculated isoelectric points of chicken and horse ferritin subunits. By two-dimensional gel electrophoresis, chicken ferritin 22 kDa subunits exhibited isoelectric points from 6.1 to 6.6 whereas horse spleen ferritin subunits exhibited isoelectric points of 5.8-6.3. The 240 kDa form of the chicken ferritin molecule had an isoelectric point of 6.6 whereas the 210 kDa form of the horse ferritin molecule had isoelectric points of 5.1 and 4.9. Intact chicken liver ferritin particles were 13.4 +/- 0.8 nm (controlled-pore glass-purified) and 12.5 +/- 0.9 nm (affinity-purified) in diameter when viewed by electron microscopy. Horse spleen ferritin consisted of slightly smaller particles with an average diameter of 11.0 +/- 0.7 nm. However, ferritin from chicken liver and horse spleen co-migrated with an apparent molecular mass of 470 kDa when analysed by Sepharose 4B gel filtration chromatography. These results indicate that, consistent with results from other published purification methods, the chicken ferritin purified by the methods reported here exhibits both structural similarities to, and differences from, horse spleen ferritin.

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Electrophoresis of Ferritins

Cited by 13 publications

References 4 publications

Mechanisms for the formation of ferritin oligomers

Mechanisms for the formation of ferritin oligomers

Histamine receptors of the microvascular endothelium revealed in situ with a histamine-ferritin conjugate: characteristic high-affinity binding sites in venules.

Purification of chicken liver ferritin by two novel methods and structural comparison with horse spleen ferritin

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