Selective Isolation of the Carboxy Terminal Peptide from a Protein

  • Tomoko Hayashi Akiyama
  • Tatsuru Sasagawa
Part of the Methods in Molecular Biology™ book series (MIMB, volume 64)


In contrast to N-termmal sequencing, several methods have been used for determination of the C-terminus. They include a thiocyanate degradation procedure (1), methods using carboxypeptidase (2), hydrazinolysis (3), and tritium labeling (4), depending on the nature of the samples.


Sodium Acetate Aromatic Amino Acid Ammonium Bicarbonate Terminal Amino Acid Selective Isolation 
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  1. 1.
    Bailey, J. M., Tu, O., Issai, G., Ha, A., and Shively, J. E. (1995) Automated carboxy-terminal sequence analysis of polypeptides containing C-terminal proline. Anal. Biochem. 224, 588–596.CrossRefGoogle Scholar
  2. 2.
    Zuber, H. and Hayashi, R. (1976) Carboxy peptidase C, carboxy peptidase Y. Methods Enzymol. 45, 561–587.CrossRefGoogle Scholar
  3. 3.
    Akabori, S., Ohno, K, and Narita, K. (1952) On the hydrazinolysis of proteins and peptides; a method for the characterization of carboxy-terminal amino acids in protein. Bull. Chem. Sot. Jap. 25, 214.CrossRefGoogle Scholar
  4. 4.
    Matsuo, H. and Narita, K. (1975) Improved Tritium-labeling for quantitative C-terminal analysis, in Protein Sequence Determination, 2nd ed. (Needleman, S. B., ed.), Springer-Verlag, Berlin, pp. 104–112.CrossRefGoogle Scholar
  5. 5.
    Kumazaki, T., Terasawa, K., and Ishii, S. (1987) Affinity chromatography on immobilized anhydrotrypsin: general utility for selective isolation of C-terminal peptides from protease digests of protein. J. Biochem. 102, 1539–1546.Google Scholar
  6. 6.
    Hayashi, T. and Sasagawa, T. (1993) A method for identifying the carboxy terminal amino acid of a protein. Anal. Biochem. 209, 163–168.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 1997

Authors and Affiliations

  • Tomoko Hayashi Akiyama
    • 1
  • Tatsuru Sasagawa
    • 1
  1. 1.Toray Research CenterKamakura, KanagawaJapan

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