Abstract
Collagen has been isolated and purified from the pronase digest of the outer annulus fibrosus of degenerated and unaffected intervertebral discs obtained at necropsy. The mixture of different collagen types has been purified and fractionated by 2 subsequent DEAE-cellulose chromatographic procedures according to established methods. Collagen chain polymers were then isolated by molecular sieving. Their reduction with 2-mercaptoethanol followed by agarose chromatography resulted in the separation of 3 major peaks corresponding to gamma, beta, and alpha chains. Cyanogen bromide (CNBr) peptides and aminoacid analyses revealed the identity of the isolated alpha-chain as alpha 1(III). Further support for this finding was obtained by immunofluorescence. It is suggested that the presence of collagen type III in the outer annulus fibrosus may be related to intervertebral disc prolapse.
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- Adam M., Dostal C., Deyl Z. Collagen heterogeneity in systemic scleroderma and other diseases. J Clin Chem Clin Biochem. 1979 Jul;17(7):495–498. doi: 10.1515/cclm.1979.17.7.495. [DOI] [PubMed] [Google Scholar]
- Adam M., Fietzek P., Kühn K. Investigations on the reaction of metals with collagen in vivo. 2. The formation of cross-links in the collagen of lathyritic rats after gold treatment in vivo. Eur J Biochem. 1968 Feb;3(4):411–414. doi: 10.1111/j.1432-1033.1967.tb19545.x. [DOI] [PubMed] [Google Scholar]
- Allmann H., Fietzek P. P., Glanville R. W., Kühn K. The covalent structure of calf skin type III collagen. VI. The amino acid sequence of the carboxyterminal cyanogen bromide peptide alpha 1(III)CB9B (position 928--1028). Hoppe Seylers Z Physiol Chem. 1979 Jul;360(7):861–868. [PubMed] [Google Scholar]
- Aumailley M., Bricaud H. Collagen synthesis in organ culture of normal and atherosclerotic aortas. Atherosclerosis. 1981 Apr;39(1):1–9. doi: 10.1016/0021-9150(81)90082-4. [DOI] [PubMed] [Google Scholar]
- Beard H. K., Roberts S., O'Brien J. P. Immunofluorescent staining for collagen and proteoglycan in normal and scoliotic intervertebral discs. J Bone Joint Surg Br. 1981;63B(4):529–534. doi: 10.1302/0301-620X.63B4.6170646. [DOI] [PubMed] [Google Scholar]
- Bentz H., Fietzek P. P., Kühn K. The covalent structure of calf skin type III collagen. III. The amino acid sequence of the cyanogen bromide peptide alpha 1(III)CB4 (positions 403--551). Hoppe Seylers Z Physiol Chem. 1979 Jul;360(7):833–840. [PubMed] [Google Scholar]
- Bornstein P., Piez K. A. The nature of the intramolecular cross-links in collagen. The separation and characterization of peptides from the cross-link region of rat skin collagen. Biochemistry. 1966 Nov;5(11):3460–3473. doi: 10.1021/bi00875a012. [DOI] [PubMed] [Google Scholar]
- Byers P. H., McKenney K. H., Lichtenstein J. R., Martin G. R. Preparation of type III procollagen and collagen from rat skin. Biochemistry. 1974 Dec 3;13(25):5243–5248. doi: 10.1021/bi00722a030. [DOI] [PubMed] [Google Scholar]
- Dewes H., Fietzek P. P., Kühn K. The covalent structure of calf skin type III collagen. II. The amino acid sequence of the cyanogen bromide peptide alpha 1(III)CB1,8,10,2(Positions 223--402). Hoppe Seylers Z Physiol Chem. 1979 Jul;360(7):821–832. [PubMed] [Google Scholar]
- Dewes H., Fietzek P. P., Kühn K. The covalent structure of calf skin type III collagen. V. The amino acid sequence of the cyanogen bromide peptide alpha 1(III)CB9A (position 789 to 927). Hoppe Seylers Z Physiol Chem. 1979 Jul;360(7):851–860. [PubMed] [Google Scholar]
- Dixit S. N. Type-IV collagens. Isolation and characterization of two structurally distinct collagen chains from bovine kidney cortices. Eur J Biochem. 1980 May;106(2):563–570. doi: 10.1111/j.1432-1033.1980.tb04604.x. [DOI] [PubMed] [Google Scholar]
- Eyre D. R. Biochemistry of the intervertebral disc. Int Rev Connect Tissue Res. 1979;8:227–291. doi: 10.1016/b978-0-12-363708-6.50012-6. [DOI] [PubMed] [Google Scholar]
- Eyre D. R., Muir H. Collagen polymorphism: two molecular species in pig intervertebral disc. FEBS Lett. 1974 Jun 1;42(2):192–196. doi: 10.1016/0014-5793(74)80783-0. [DOI] [PubMed] [Google Scholar]
- Eyre D. R., Muir H. Quantitative analysis of types I and II collagens in human intervertebral discs at various ages. Biochim Biophys Acta. 1977 May 27;492(1):29–42. doi: 10.1016/0005-2795(77)90211-2. [DOI] [PubMed] [Google Scholar]
- Eyre D. R., Muir H. Types I and II collagens in intervertebral disc. Interchanging radial distributions in annulus fibrosus. Biochem J. 1976 Jul 1;157(1):267–270. doi: 10.1042/bj1570267. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Furthmayr H., Timpl R. Characterization of collagen peptides by sodium dodecylsulfate-polyacrylamide electrophoresis. Anal Biochem. 1971 Jun;41(2):510–516. doi: 10.1016/0003-2697(71)90173-4. [DOI] [PubMed] [Google Scholar]
- Happey F., Pearson C. H., Naylor A., Turner R. L. The ageing of the human intervertebral disc. Gerontologia. 1969;15(2):174–188. doi: 10.1159/000211685. [DOI] [PubMed] [Google Scholar]
- Lang H., Glanville R. W., Fietzek P. P., Kühn K. The covalent structure of calf skin type III collagen. IV. The amino acid sequence of the cyanogen bromide peptide alpha 1(III)CB5 (positions 552--788). Hoppe Seylers Z Physiol Chem. 1979 Jul;360(7):841–850. [PubMed] [Google Scholar]
- Miller E. J. Isolation and characterization of a collagen from chick cartilage containing three identical alpha chains. Biochemistry. 1971 Apr 27;10(9):1652–1659. doi: 10.1021/bi00785a024. [DOI] [PubMed] [Google Scholar]
- Nowack H., Gay S., Wick G., Becker U., Timpl R. Preparation and use in immunohistology of antibodies specific for type I and type III collagen and procollagen. J Immunol Methods. 1976;12(1-2):117–124. doi: 10.1016/0022-1759(76)90101-0. [DOI] [PubMed] [Google Scholar]
- Piez K. A. Molecular weight determination of random coil polypeptides from collagen by molecular sieve chromatography. Anal Biochem. 1968 Nov;26(2):305–312. doi: 10.1016/0003-2697(68)90342-4. [DOI] [PubMed] [Google Scholar]
- Timpl R., Glanville R. W., Wick G., Martin G. R. Immunochemical study on basement membrane (type IV) collagens. Immunology. 1979 Sep;38(1):109–116. [PMC free article] [PubMed] [Google Scholar]
- von der Mark H., von der Mark K., Gay S. Study of differential collagen synthesis during development of the chick embryo by immunofluorescence. I. Preparation of collagen type I and type II specific antibodies and their application to early stages of the chick embryo. Dev Biol. 1976 Feb;48(2):237–249. doi: 10.1016/0012-1606(76)90088-9. [DOI] [PubMed] [Google Scholar]