Physical, chemical, and ultrastructural studies of water-soluble human amyloid fibrils. Comparative analyses of nine amyloid preparations.
Amyloid fibrils were isolated from the tissues of nine patients with amyloidosis in a state of high purity by homogenization of the tissue followed by extraction with distilled water. Physical, chemical, and ultrastructural studies suggest that amyloid fibrils from different individuals resemble each other, but are not identical. In tissue sections as well as by negative staining of isolated fibrils, morphologic variations were observed. Among the isolated fibrils at least three types were noted. The majority resembled those described previously. However, one subject had two types of fibrils which differed in size and appearance. Most of the preparations sedimented as a single component with a sedimentation coefficient of 45-50S or as a larger polymer. However, two of the preparations had sedimentation coefficients of 8-9S, and a third one had a major 95S component and a minor 9S fraction. While the preparations of amyloid were not sufficiently pure for amino acid analyses, peptide maps demonstrated differences among amyloid preparations from different subjects. The amyloid fibrils in their native state proved to be remarkably resistant to digestion by a number of proteolytic enzymes. Several chemical methods were tried to produce smaller subunits. Of these, the most successful one was the use of 0.1 M NaOH which yielded a smaller, soluble fraction with sedimentation coefficients ranging from 1.1 to 2.8S. Accompanying this degradation, there was little loss of peptides or carbohydrates. Based on the results of the chemical analyses, it is estimated that the subunit produced by sodium hydroxide had a molecular weight of approximately 35,000-40,000.
D000293:Adolescent; D000368:Aged; D000596:Amino Acids / Q000032:analysis; D000682:Amyloid / Q000032:analysis* / Q000145:classification; D055598:Chemical Phenomena; D002621:Chemistry; D002627:Chemistry, Physical; D003224:Congo Red; D005260:Female; D006321:Heart / Q000032:analysis; D006801:Humans; D007668:Kidney / Q000032:analysis; D008099:Liver / Q000032:analysis; D008198:Lymph Nodes / Q000032:analysis; D008297:Male; D008854:Microscopy, Electron; D009101:Multiple Myeloma / Q000378:metabolism; D010455:Peptides / Q000032:analysis; D012995:Solubility; D013154:Spleen / Q000032:analysis; D014059:Tongue / Q000032:analysis; D014461:Ultracentrifugation
D000596:Amino Acids; D000682:Amyloid; D010455:Peptides; D003224:Congo Red