Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1980 Oct 1;191(1):253–256. doi: 10.1042/bj1910253

Separation of collagen cyanogen bromide-derived peptides by reversed-phase high-performance liquid chromatography.

M van der Rest, H P Bennett, S Solomon, F H Glorieux
PMCID: PMC1162205  PMID: 7470094

Abstract

A new technique for separation of the CNBr cleavage products of collagen is described. It involves the use of a 30 nm pore reversed-phase high-performance liquid chromatography column eluted with a linear gradient of acetonitrile/water containing 0.01 M-heptafluorobutyric acid. The separation of type I, type II and type III CNBr peptides is described. Resolution is particularly good for the low-molecular-weight peptides. The method is fast, quantitative and sensitive, and the complete volatility of the eluent facilitates the recovery of the separated peptides.

Full text

PDF
253

Selected References

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

  1. 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]
  2. Butler W. T., Piez K. A., Bornstein P. Isolation and characterization of the cyanogen bromide peptides from the alpha-1 chain of rat skin collagen. Biochemistry. 1967 Dec;6(12):3771–3780. doi: 10.1021/bi00864a022. [DOI] [PubMed] [Google Scholar]
  3. Cole W. G., Bean D. A. Analysis of collagen cyanogen bromide peptides using electrophoresis in continuous concave gradient polyacrylamide gels. Anal Biochem. 1979 Jan 1;92(1):183–188. doi: 10.1016/0003-2697(79)90642-0. [DOI] [PubMed] [Google Scholar]
  4. Eyre D. R., Muir H. Characterisation of the major CNBr-Derived peptides of porcine type II collagen. Connect Tissue Res. 1975;3(2):165–170. doi: 10.3109/03008207509152175. [DOI] [PubMed] [Google Scholar]
  5. Fietzek P. P., Allmann H., Rauterberg J., Wachter E. Ordering of cyanogen bromide peptides of type III collagen based on their homology to type I collagen: preservation of sites for crosslink formation during evolution. Proc Natl Acad Sci U S A. 1977 Jan;74(1):84–86. doi: 10.1073/pnas.74.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fietzek P. P., Kühn K. The primary structure of collagen. Int Rev Connect Tissue Res. 1976;7:1–60. doi: 10.1016/b978-0-12-363707-9.50007-1. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Piez K. A., Sherman M. R. Characterization of the product formed by renaturation of alpha 1-CB2, a small peptide from collagen. Biochemistry. 1970 Oct 13;9(21):4129–4133. doi: 10.1021/bi00823a015. [DOI] [PubMed] [Google Scholar]
  9. Piez K. A., Sherman M. R. Equilibrium and kinetic studies of the helix-coil transition in alpha 1-CB2, a small peptide from collagen. Biochemistry. 1970 Oct 13;9(21):4134–4140. doi: 10.1021/bi00823a016. [DOI] [PubMed] [Google Scholar]
  10. Scott P. G., Veis A. The cyanogen bromide peptides of bovine soluble and insoluble collagens. I. Characterization of peptides from soluble type I collagen by sodium dodecylsulphate polyacrylamide gel electrophoresis. Connect Tissue Res. 1976;4(2):107–116. doi: 10.3109/03008207609152206. [DOI] [PubMed] [Google Scholar]
  11. van Der Rest M., Cole W. G., Glorieux F. H. Human collagen 'fingerprints' produced by clostridopeptidase A digestion and high-pressure liquid chromatography. Biochem J. 1977 Mar 1;161(3):527–534. doi: 10.1042/bj1610527. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES