Advertisement

Automated C-Terminal Sequencing of Polypeptides Containing C-Terminal Proline

  • Jerome M. Bailey
  • Oanh Tu
  • Gilbert Issai
  • John E. Shively

Abstract

There has been much interest in the development of a chemical method for the sequential C-terminal sequence analysis of proteins and peptides. Such a method would be analogous and complimentary to the Edman degradation commonly used for N-terminal sequence analysis (Edman, 1950). It would also be invaluable for the sequence analysis of proteins with naturally occurring N-terminal blocking groups, for the detection of post-translational processing at the carboxy-terminus of expressed gene products, and for assistance in the design of oligonucleotide probes for gene cloning. Although a number of methods have been described, the method known as the “thiocyanate method” (Schlack and Kumpf, 1926), has been the most widely studied and appears to offer the most promise due to its similarity to current methods of N-terminal sequence analysis.

Keywords

Trifluoroacetic Acid Acetic Anhydride Quaternary Amine Ammonium Thiocyanate Common Amino Acid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bailey, J.M., and Shively, J.E. (1990) Carboxy-terminal sequencing: Formation and hydrolysis of C-terminal peptidylthiohydantoins, Biochemistry 29, 3145–3156.PubMedCrossRefGoogle Scholar
  2. Bailey, J.M, Shenoy, N.S., Ronk, M., and Shively, J.E. (1992a) Automated carboxy-terminal sequence analysis of peptides, Protein Science 1, 68–80.PubMedCrossRefGoogle Scholar
  3. Bailey, J.M., Nikfarjam, E, Shenoy, N.S., and Shively, J.E. (1992b) Automated Carboxy-Terminal Sequence Analysis of Peptides and Proteins Using Diphenyl Phosphoroisothio-cyanatidate, Protein Science 1, 1622–1633.PubMedCrossRefGoogle Scholar
  4. Bailey, J.M., Rusnak, M., and Shively, J.E. (1993) Analytical Biochemistry 212, 366–374.Google Scholar
  5. Edman, P. (1950) Method for Determination of the Amino Acid Sequence in Peptides, Acta Chem. Scand. 4, 283–293.CrossRefGoogle Scholar
  6. Fox, S.W., Hurst, T.L., Griffith, J. F., and Underwood, 0. (1955) A method for the quantitative determination of C-terminal amino acid residues, J. Am. Chem. Soc. 77, 3119–3122.CrossRefGoogle Scholar
  7. Inglis, A.S., Wilshire, J.F.K., Casagranda, F., and Laslett, R.L. (1989) C-Terminal sequencing: A new look at the Schlack-Kumpf thiocyanate degradation, in Methods in Protein Sequence Analysis (WittmannLiebold, B., Ed.) pp.137–144, Springer-Verlag.CrossRefGoogle Scholar
  8. Inglis, A.S., and De Luca, C. (1993) A new chemical approach to C-terminal microsequence analysis via the thiohydantoin, in Methods in Protein Sequence Analysis (Imahori, K.,Sakiyama, F., Eds.) 71–78, Plenum Publishing Corp.Google Scholar
  9. Kenner, G.W., Khorana, H.G., and Stedman, R.J. (1953) Peptides. Part IV. Selective removal of the C-terminal Residue as a thiohydantoin. The use of diphenylphosphoroisothiocyanatidate, Chem. Soc. Jour. (London), 673–678.Google Scholar
  10. Kubo, H., Nakajima, T., and Tamura, Z. (1971) Formation of thiohydantoin derivative of proline from the C-terminal of peptides, Chem. Pharm. Bull. 19, 210–211.CrossRefGoogle Scholar
  11. Schlack, P., and Kumpf, W. (1926) Uber eine neue methode zur Ermittlung der konstitution von peptiden, Z. Physiol. Chem. 154, 125–170.CrossRefGoogle Scholar
  12. Shenoy, N.S., Bailey, J.M., and Shively, J.E. (1992) Carboxylic acid modified polyethylene: A novel support for the covalent immobilization of polypeptides for C-terminal sequencing, Protein Science 1, 58–67.PubMedCrossRefGoogle Scholar
  13. Stark, G.R. (1968) Sequential degradation of peptides from their carboxyl termini with ammonium thiocyanate and acetic anhydride, Biochem. 7, 1796–1807.CrossRefGoogle Scholar
  14. Turner, R.A., and Schmerzler, G. (1954) Identification of C-terminal residues in peptides and proteins through formation of thiohydantoins, Biochim. Biophys. Acta. 13, 553–559.PubMedCrossRefGoogle Scholar
  15. Yamashita, S., and Ishikawa, N. (1971) Sequential degradation of polypeptides from the carboxyl ends II. Application to polypeptides, Proc. Hoshi. Pharm. 13, 136–138.Google Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Jerome M. Bailey
    • 1
  • Oanh Tu
    • 1
  • Gilbert Issai
    • 1
  • John E. Shively
    • 1
  1. 1.Division of ImmunologyBeckman Research Institute of the City of HopeDuarteUSA

Personalised recommendations