[go: up one dir, main page]

Skip to main content

Advertisement

Log in

Isolation of peripheral nerve collagen

  • Original Articles
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

A new method for the isolation of high yields of collagen from human peripheral nerve is described. A major technique adopted in the present work is sedimentation of the tissue homogenates in a sucrose density gradient. The defatted fibrous material isolated after the removal of myelin was shown to be a relatively pure collagenous substance by amino acid analysis, indicating that the removal of noncollagenous proteins, especially glycoproteins, from collagen fibrils was effectively achieved by this method. The yield of collagen at this step was more than 90% of the total collagen in peripheral nerve. Subsequent extractions with solutions of neutral saline and sodium citrate were found to give further purification of the collagenous protein. The collagens from embryonic peripheral nerves were composed of Type I and III collagens, while Type III collagen was found to be less abundant in adult peripheral nerves.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Thomas, P. K. 1963. The connective tissue of peripheral nerve: an electron microscope study. J. Anat. 97:35–44.

    PubMed  Google Scholar 

  2. Shellswell, G. B., Restall, D. J., Duance, V. C., andBailey, A. J. 1979. Identification and differential distribution of collagen types in the central and peripheral nervous systems. FEBS Letters 106:305–308.

    PubMed  Google Scholar 

  3. Koeppen, A. H., Messmore, H., andStehbens, W. E. 1971. Interstitial hypertrophic neuropathy. Biochemical study of the periphral nervous system. Arch. Neurol. 24:340–352.

    PubMed  Google Scholar 

  4. Pleasure, D., Bora, F. W., andLane, J. 1974. Regeneration after nerve transection: Effect of inhibition of collagen synthesis. Exp. Neurol. 45:72–78.

    PubMed  Google Scholar 

  5. Seyer, J. M., Kang, A. H., andWhitaker, J. N. 1977. The characterization of Type I and Type III collagens from human peripheral nerve. Biochim. Biophys. Acta 492:415–425.

    PubMed  Google Scholar 

  6. Everly, J. L., Brady, R. O., andQuarles, R. H. 1973. Evidence that major protein in rat sciatic nerve myelin is a glycoprotein. J. Neurochem. 21:329–334.

    PubMed  Google Scholar 

  7. Piez, K., Eigner, E. A., andLewis, M. S. 1963. The chromatographic separation and amino acid composition of the subunits of several collagens. Biochemistry 2:58–66.

    Google Scholar 

  8. Fujii, K., Kajiwara, T., andKurosu, H. 1979. Effect of vitamin B6 deficiency on the crosslink formation of collagen. FEBS Letters97:193–195.

    PubMed  Google Scholar 

  9. Bergman, I., andLoxley, R. 1963. Two improved and simplified methods for the spectrophotometric determination of hydroxyproline. Anal. Chem. 35:1961–1965.

    Google Scholar 

  10. Rubin, A. L., Drake, M. P., Davison, P. E., Pfahl, D., Speakman, P. T., andSchmitt, F. O. 1965. Effects of pepsin treatment on the interaction properties of tropocollagen macromolecules. Biochemistry 4:181–190.

    Google Scholar 

  11. Sykes, B., Puddle, B., Francis, M., andSmith, R. 1976. The estimation of two collagens from human dermis by interrupted gel electrophoresis. Biochem. Biophys. Res. Commun. 72:1472–1480.

    PubMed  Google Scholar 

  12. Fujii, K., Tanzer, M. L., Nusgens, B. V., andLapiere, C. M. 1975. Aldehyde content and cross-linking of Type III collagen. Biochem. Biophys. Res. Commun. 69:128–134.

    Google Scholar 

  13. Miller, E. J., Epstein, E. H., andPiez, K. A. 1971. Identification of three genetically distinct collagens by cyanogen bromide cleavege of insoluble human skin and cartilage collagen. Biochem. Biophys. Res. Commun. 42:1024–1029.

    PubMed  Google Scholar 

  14. Timpl, R., Wick, G., andGray, S. 1977. Antibodies to distinct types of collagens and procollagens and their application in immunohistology. J. Immunol. Methods 18:165–182.

    PubMed  Google Scholar 

  15. Hahn, E., andTimpl, R. 1973. Involvement of more than a single polypeptide chain in the helical antigenic determinants of collagen. Eur. J. Immunol. 3:442–446.

    PubMed  Google Scholar 

  16. March, S., Parikh, I., andCautrecasa, P. 1974. A simplified method for cyanogen bromide activation of Agarose for affinity chromatography. Anal. Biochem. 60:149–152.

    PubMed  Google Scholar 

  17. Epstein, E. H. Jr. 1974. [αl(III)]3 human skin collagen. Release by pepsin digestion and preponderance in fetal life. J. Biol. Chem. 249:3225–3231.

    PubMed  Google Scholar 

  18. Shuttleworth, C. A., andForrest, L. 1975. Changes in guinea-pig dermal collagen during development. Eur. J. Biochem. 55:391–395.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fujii, K., Tsuji, M. & Murota, K. Isolation of peripheral nerve collagen. Neurochem Res 11, 1439–1446 (1986). https://doi.org/10.1007/BF00966223

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00966223

Keywords

Navigation