Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1987 Dec 1;248(2):373–381. doi: 10.1042/bj2480373

Type VI collagen of the intervertebral disc. Biochemical and electron-microscopic characterization of the native protein.

J J Wu 1, D R Eyre 1, H S Slayter 1
PMCID: PMC1148551  PMID: 3124811

Abstract

The collagen framework of the intervertebral disc contains two major fibril-forming collagens, types I and II. Smaller amounts of other types of collagen are also present. On examination of the nature and distribution of these minor collagens within bovine disc tissue, type VI collagen was found to be unusually abundant. It accounted for about 20% of the total collagen in calf nucleus pulposus, and about 5% in the annulus fibrosus. It was discovered by serially digesting disc tissue with chondroitin ABC lyase and Streptomyces hyaluronidase that native covalent polymers of type VI collagen could be extracted. Electron micrographs of this material prepared by rotary shadowing revealed the characteristic dimensions of tetramers and double tetramers of type VI molecules, with their central rods and terminal globular domains. Molecular-sieve column chromatography on agarose under non-reducing non-denaturing conditions gave a series of protein peaks with molecular sizes equivalent to the tetramer, double tetramer and higher multimers. On SDS/polyacrylamide-gel electrophoresis after disulphide cleavage, these fractions of type VI collagen all showed a main band at Mr 140,000 and four lesser bands between Mr 180,000 and 240,000. On electrophoresis without disulphide cleavage in agarose/2.4% polyacrylamide only dimeric (six chains) and tetrameric (12 chains) forms of type VI molecules were present. The ability to extract all the type VI collagen of the tissue in 4 M-guanidinium chloride, and absence of aldehyde-mediated cross-linking residues on direct analysis, showed that, in contrast with most matrix collagens, type VI collagen does not function as a covalently cross-linked structural polymer.

Full text

PDF
373

Images in this article

375Fig. 1.
on p.375
375Fig. 2.
on p.375
376Fig. 3.
on p.376
377Fig. 4.
on p.377
378Fig. 5.
on p.378
379Fig. 6.
on p.379

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abedin M. Z., Ayad S., Weiss J. B. Isolation and native characterization of cysteine-rich collagens from bovine placental tissues and uterus and their relationship to types IV and V collagens. Biosci Rep. 1982 Jul;2(7):493–502. doi: 10.1007/BF01115247. [DOI] [PubMed] [Google Scholar]
  2. Ayad S., Abedin M. Z., Grundy S. M., Weiss J. B. Isolation and characterisation of an unusual collagen from hyaline cartilage and intervertebral disc. FEBS Lett. 1981 Jan 26;123(2):195–199. doi: 10.1016/0014-5793(81)80286-4. [DOI] [PubMed] [Google Scholar]
  3. Ayad S., Abedin M. Z., Weiss J. B., Grundy S. M. Characterisation of another short-chain disulphide-bonded collagen from cartilage, vitreous and intervertebral disc. FEBS Lett. 1982 Mar 22;139(2):300–304. doi: 10.1016/0014-5793(82)80875-2. [DOI] [PubMed] [Google Scholar]
  4. Ayad S., Chambers C. A., Berry L., Shuttleworth C. A., Grant M. E. Type VI collagen and glycoprotein MFPI are distinct components of the extracellular matrix. Biochem J. 1986 May 15;236(1):299–302. doi: 10.1042/bj2360299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ayad S., Chambers C. A., Shuttleworth C. A., Grant M. E. Isolation from bovine elastic tissues of collagen type VI and characterization of its form in vivo. Biochem J. 1985 Sep 1;230(2):465–474. doi: 10.1042/bj2300465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bruns R. R. Beaded filaments and long-spacing fibrils: relation to type VI collagen. J Ultrastruct Res. 1984 Nov;89(2):136–145. doi: 10.1016/s0022-5320(84)80010-6. [DOI] [PubMed] [Google Scholar]
  7. Bruns R. R., Press W., Engvall E., Timpl R., Gross J. Type VI collagen in extracellular, 100-nm periodic filaments and fibrils: identification by immunoelectron microscopy. J Cell Biol. 1986 Aug;103(2):393–404. doi: 10.1083/jcb.103.2.393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Buckwalter J. A., Maynard J. A., Cooper R. R. Banded structures in human nucleus pulposus. Clin Orthop Relat Res. 1979 Mar-Apr;(139):259–266. [PubMed] [Google Scholar]
  9. Chung E., Rhodes K., Miller E. J. Isolation of three collagenous components of probable basement membrane origin from several tissues. Biochem Biophys Res Commun. 1976 Aug 23;71(4):1167–1174. doi: 10.1016/0006-291x(76)90776-2. [DOI] [PubMed] [Google Scholar]
  10. Cornah M. S., Meachim G., Parry E. W. Banded structures in the matrix of human and rabbit nucleus pulposus. J Anat. 1970 Sep;107(Pt 2):351–362. [PMC free article] [PubMed] [Google Scholar]
  11. Engel J., Furthmayr H., Odermatt E., von der Mark H., Aumailley M., Fleischmajer R., Timpl R. Structure and macromolecular organization of type VI collagen. Ann N Y Acad Sci. 1985;460:25–37. doi: 10.1111/j.1749-6632.1985.tb51154.x. [DOI] [PubMed] [Google Scholar]
  12. Engvall E., Hessle H., Klier G. Molecular assembly, secretion, and matrix deposition of type VI collagen. J Cell Biol. 1986 Mar;102(3):703–710. doi: 10.1083/jcb.102.3.703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Eyre D. R., Koob T. J., Van Ness K. P. Quantitation of hydroxypyridinium crosslinks in collagen by high-performance liquid chromatography. Anal Biochem. 1984 Mar;137(2):380–388. doi: 10.1016/0003-2697(84)90101-5. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Eyre D. R., Oguchi H. The hydroxypyridinium crosslinks of skeletal collagens: their measurement, properties and a proposed pathway of formation. Biochem Biophys Res Commun. 1980 Jan 29;92(2):403–410. doi: 10.1016/0006-291x(80)90347-2. [DOI] [PubMed] [Google Scholar]
  17. Eyre D. R., Paz M. A., Gallop P. M. Cross-linking in collagen and elastin. Annu Rev Biochem. 1984;53:717–748. doi: 10.1146/annurev.bi.53.070184.003441. [DOI] [PubMed] [Google Scholar]
  18. Eyre D. R., Wu J. J. Collagen of fibrocartilage: a distinctive molecular phenotype in bovine meniscus. FEBS Lett. 1983 Jul 25;158(2):265–270. doi: 10.1016/0014-5793(83)80592-4. [DOI] [PubMed] [Google Scholar]
  19. Eyre D. R., Wu J. J., Woolley D. E. All three chains of 1 alpha 2 alpha 3 alpha collagen from hyaline cartilage resist human collagenase. Biochem Biophys Res Commun. 1984 Feb 14;118(3):724–729. doi: 10.1016/0006-291x(84)91454-2. [DOI] [PubMed] [Google Scholar]
  20. Eyre D. Collagen cross-linking amino acids. Methods Enzymol. 1987;144:115–139. doi: 10.1016/0076-6879(87)44176-1. [DOI] [PubMed] [Google Scholar]
  21. Furthmayr H., Wiedemann H., Timpl R., Odermatt E., Engel J. Electron-microscopical approach to a structural model of intima collagen. Biochem J. 1983 May 1;211(2):303–311. doi: 10.1042/bj2110303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Furuto D. K., Miller E. J. Characterization of a unique collagenous fraction from limited pepsin digests of human placental tissue: molecular organization of the native aggregate. Biochemistry. 1981 Mar 17;20(6):1635–1640. doi: 10.1021/bi00509a035. [DOI] [PubMed] [Google Scholar]
  23. Furuto D. K., Miller E. J. Isolation of a unique collagenous fraction from limited pepsin digests of human placental tissue. Characterization of one of the constituent polypeptide chains. J Biol Chem. 1980 Jan 10;255(1):290–295. [PubMed] [Google Scholar]
  24. Hessle H., Engvall E. Type VI collagen. Studies on its localization, structure, and biosynthetic form with monoclonal antibodies. J Biol Chem. 1984 Mar 25;259(6):3955–3961. [PubMed] [Google Scholar]
  25. Jander R., Rauterberg J., Glanville R. W. Further characterization of the three polypeptide chains of bovine and human short-chain collagen (intima collagen). Eur J Biochem. 1983 Jun 1;133(1):39–46. doi: 10.1111/j.1432-1033.1983.tb07427.x. [DOI] [PubMed] [Google Scholar]
  26. Jander R., Rauterberg J., Voss B., von Bassewitz D. B. A cysteine-rich collagenous protein from bovine placenta. Isolation of its constituent polypeptide chains and some properties of the non-denatured protein. Eur J Biochem. 1981;114(1):17–25. [PubMed] [Google Scholar]
  27. Jander R., Troyer D., Rauterberg J. A collagen-like glycoprotein of the extracellular matrix is the undegraded form of type VI collagen. Biochemistry. 1984 Jul 31;23(16):3675–3681. doi: 10.1021/bi00311a016. [DOI] [PubMed] [Google Scholar]
  28. Knight K. R., Ayad S., Shuttleworth C. A., Grant M. E. A collagenous glycoprotein found in dissociative extracts of foetal bovine nuchal ligament. Evidence for a relationship with type VI collagen. Biochem J. 1984 Jun 1;220(2):395–403. doi: 10.1042/bj2200395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  30. Miller E. J. The structure of fibril-forming collagens. Ann N Y Acad Sci. 1985;460:1–13. doi: 10.1111/j.1749-6632.1985.tb51152.x. [DOI] [PubMed] [Google Scholar]
  31. Odermatt E., Risteli J., van Delden V., Timpl R. Structural diversity and domain composition of a unique collagenous fragment (intima collagen) obtained from human placenta. Biochem J. 1983 May 1;211(2):295–302. doi: 10.1042/bj2110295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Peacock A. C., Dingman C. W. Molecular weight estimation and separation of ribonucleic acid by electrophoresis in agarose-acrylamide composite gels. Biochemistry. 1968 Feb;7(2):668–674. doi: 10.1021/bi00842a023. [DOI] [PubMed] [Google Scholar]
  33. Slayter H. S. High-resolution metal replication of macromolecules. Ultramicroscopy. 1976 Sep-Oct;1(4):341–357. doi: 10.1016/0304-3991(76)90050-4. [DOI] [PubMed] [Google Scholar]
  34. Slayter H., Loscalzo J., Bockenstedt P., Handin R. I. Native conformation of human von Willebrand protein. Analysis by electron microscopy and quasi-elastic light scattering. J Biol Chem. 1985 Jul 15;260(14):8559–8563. [PubMed] [Google Scholar]
  35. Trüeb B., Bornstein P. Characterization of the precursor form of type VI collagen. J Biol Chem. 1984 Jul 10;259(13):8597–8604. [PubMed] [Google Scholar]
  36. Wu J. J., Eyre D. R. Cartilage type IX collagen is cross-linked by hydroxypyridinium residues. Biochem Biophys Res Commun. 1984 Sep 28;123(3):1033–1039. doi: 10.1016/s0006-291x(84)80237-5. [DOI] [PubMed] [Google Scholar]
  37. Zimmermann D. R., Trüeb B., Winterhalter K. H., Witmer R., Fischer R. W. Type VI collagen is a major component of the human cornea. FEBS Lett. 1986 Mar 3;197(1-2):55–58. doi: 10.1016/0014-5793(86)80297-6. [DOI] [PubMed] [Google Scholar]
  38. von der Mark H., Aumailley M., Wick G., Fleischmajer R., Timpl R. Immunochemistry, genuine size and tissue localization of collagen VI. Eur J Biochem. 1984 Aug 1;142(3):493–502. doi: 10.1111/j.1432-1033.1984.tb08313.x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES