Structural heterogeneity and subunit composition of horse ferritins.
Structural, spectroscopic, and immunological properties of horse ferritins extracted from spleen, liver, and heart were studied to test the hypothesis that the different tissue ferritins are hybrids composed of variable proportions of two subunit types. The weight-average molecular weights determined by sedimentation velocity and gel filtration increase from 460 000 for spleen to 480 000 for liver and to 515 000 for heart apoferritin; moreover, the diffusion coefficients prove that each tissue-specific ferritin consists of a population of hybrid molecules. The intrinsic fluorescence and the near-UV circular dichroism (CD) spectra change as a function of the subunit composition of the three ferritins. The fluorescence emission maximum, which occurs at a very low wavelength (315 nm) in horse spleen apoferritin, is shifted to increasingly higher wavelengths in the liver and heart proteins (320 and 325 nm, respectively), indicating that the tryptophan and tyrosine residues become less rigidly immobilized with an increase in the H-subunit content. The tryptophan residues behave as fully solvated in the monomeric subunits at acidic pH values. In accordance with the fluorescence data, the near-UV CD spectra show that the tryptophan environment is highly asymmetric in spleen apoferritin, progressively less so in liver and heart apoferritins, and completely relaxed in the dissociated subunits. Moreover, they show that the environment of tyrosines and phenylalanines differs markedly in the spleen and heart apoproteins. The ferritins studied appear to have immunogenic sites which are specific for the H and L subunits on the basis of enzyme-linked immunoassay and double-diffusion experiments.
D000818:Animals; D001052:Apoferritins / Q000276:immunology; D055598:Chemical Phenomena; D002621:Chemistry; D005293:Ferritins / Q000276:immunology; D006736:Horses; D007120:Immunochemistry; D008099:Liver / Q000032:analysis; D008970:Molecular Weight; D009206:Myocardium / Q000032:analysis; D009928:Organ Specificity; D011487:Protein Conformation; D013154:Spleen / Q000032:analysis
D005293:Ferritins; D001052:Apoferritins