Abstract
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.
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- BRENNER S., HORNE R. W. A negative staining method for high resolution electron microscopy of viruses. Biochim Biophys Acta. 1959 Jul;34:103–110. doi: 10.1016/0006-3002(59)90237-9. [DOI] [PubMed] [Google Scholar]
- Cathcart E. S., Wollheim F. A., Cohen A. S. Plasma protein constituents of amyloid fibrils. J Immunol. 1967 Aug;99(2):376–385. [PubMed] [Google Scholar]
- Cohen A. S. Amyloidosis. N Engl J Med. 1967 Sep 7;277(10):522–contd. doi: 10.1056/NEJM196709072771006. [DOI] [PubMed] [Google Scholar]
- Franklin E. C., Pras M. Immunologic studies of water-soluble human amyloid fibrils. Comparative studies of eight amyloid preparations. J Exp Med. 1969 Oct 1;130(4):797–808. doi: 10.1084/jem.130.4.797. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GUEFT B., GHIDONI J. J. THE SITE OF FORMATION AND ULTRASTRUCTURE OF AMYLOID. Am J Pathol. 1963 Nov;43:837–854. [PMC free article] [PubMed] [Google Scholar]
- Glenner G. G., Keiser H. R., Bladen H. A., Cuatrecasas P., Eanes E. D., Ram J. S., Kanfer J. N., DeLellis R. A. Amyloid. VI. A comparison of two morphologic components of human amyloid deposits. J Histochem Cytochem. 1968 Oct;16(10):633–644. doi: 10.1177/16.10.633. [DOI] [PubMed] [Google Scholar]
- HEEFNER W. A., SORENSON G. D. Experimental amyloidosis. I. Light and electron microscopic observation of spleen and lymph nodes. Lab Invest. 1962 Aug;11:585–593. [PubMed] [Google Scholar]
- HELLER H., MISSMAHL H. P., SOHAR E., GAFNI J. AMYLOIDOSIS: ITS DIFFERENTIATION INTO PERIRETICULIN AND PERI-COLLAGEN TYPES. J Pathol Bacteriol. 1964 Jul;88:15–34. doi: 10.1002/path.1700880103. [DOI] [PubMed] [Google Scholar]
- KATZ A. M., DREYER W. J., ANFINSEN C. B. Peptide separation by two-dimensional chromatography and electrophoresis. J Biol Chem. 1959 Nov;234:2897–2900. [PubMed] [Google Scholar]
- LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MELTZER M., FRANKLIN E. C., FUNDENBERG H., FRANGIONE B. SINGLE PEPTIDE DIFFERENCES BETWEEN GAMMA-GLOBULINS OF DIFFERENT GENETIC (GM) TYPES. Proc Natl Acad Sci U S A. 1964 Jun;51:1007–1014. doi: 10.1073/pnas.51.6.1007. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MILLONIG G. A modified procedure for lead staining of thin sections. J Biophys Biochem Cytol. 1961 Dec;11:736–739. doi: 10.1083/jcb.11.3.736. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miller H. I., Rotman Y., Ben-Shaul Y., Ashkenazi Y. The dissociation of the amyloid filament so subunits. Isr J Med Sci. 1968 Sep-Oct;4(5):982–986. [PubMed] [Google Scholar]
- Pras M., Schubert M. Metachromatic properties of amyloid in solution. J Histochem Cytochem. 1969 Apr;17(4):258–265. doi: 10.1177/17.4.258. [DOI] [PubMed] [Google Scholar]
- Pras M., Schubert M., Zucker-Franklin D., Rimon A., Franklin E. C. The characterization of soluble amyloid prepared in water. J Clin Invest. 1968 Apr;47(4):924–933. doi: 10.1172/JCI105784. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reimann H. A., Koucky R. F., Eklund C. M. Primary Amyloidosis Limited to Tissue of Mesodermal Origin. Am J Pathol. 1935 Nov;11(6):977–988.3. [PMC free article] [PubMed] [Google Scholar]
- SABATINI D. D., BENSCH K., BARRNETT R. J. Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J Cell Biol. 1963 Apr;17:19–58. doi: 10.1083/jcb.17.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SPIRO D. The structural basis of proteinuria in man; electron microscopic studies of renal biopsy specimens from patients with lipid nephrosis, amyloidosis, and subacute and chronic glomerulonephritis. Am J Pathol. 1959 Jan-Feb;35(1):47–73. [PMC free article] [PubMed] [Google Scholar]
- Shirahama T., Cohen A. S. High-resolution electron microscopic analysis of the amyloid fibril. J Cell Biol. 1967 Jun;33(3):679–708. doi: 10.1083/jcb.33.3.679. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shirahama T., Cohen A. S. Reconstitution of amyloid fibrils from alkaline extracts. J Cell Biol. 1967 Nov;35(2):459–464. doi: 10.1083/jcb.35.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sri Ram J., DeLellis R. A., Glenner G. G. Amyloid. IV. Is human amyloid immunogenic? Int Arch Allergy Appl Immunol. 1968;34(3):269–282. [PubMed] [Google Scholar]
- WATSON M. L. Staining of tissue sections for electron microscopy with heavy metals. J Biophys Biochem Cytol. 1958 Jul 25;4(4):475–478. doi: 10.1083/jcb.4.4.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wright J. R., Calkins E., Breen W. J., Stolte G., Schultz R. T. Relationship of amyloid to aging. Review of the literature and systematic study of 83 patients derived rom a general hospital population. Medicine (Baltimore) 1969 Jan;48(1):39–60. [PubMed] [Google Scholar]