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Selective Isolation of the Carboxy Terminal Peptide from a Protein

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Protein Sequencing Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 64))

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Abstract

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.

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References

  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.

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  2. Zuber, H. and Hayashi, R. (1976) Carboxy peptidase C, carboxy peptidase Y. Methods Enzymol. 45, 561–587.

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  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.

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  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.

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  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.

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  6. Hayashi, T. and Sasagawa, T. (1993) A method for identifying the carboxy terminal amino acid of a protein. Anal. Biochem. 209, 163–168.

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Author information

Authors and Affiliations

  1. Toray Research Center, Kamakura, Kanagawa, Japan

    Tomoko Hayashi Akiyama & Tatsuru Sasagawa

Authors
  1. Tomoko Hayashi Akiyama
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  2. Tatsuru Sasagawa
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Editor information

Editors and Affiliations

  1. Celltech Therapeutics, Slough, UK

    Bryan John Smith

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© 1997 Humana Press Inc.

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Akiyama, T.H., Sasagawa, T. (1997). Selective Isolation of the Carboxy Terminal Peptide from a Protein. In: Smith, B.J. (eds) Protein Sequencing Protocols. Methods in Molecular Biology™, vol 64. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-353-8:81

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  • DOI: https://doi.org/10.1385/0-89603-353-8:81

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-353-5

  • Online ISBN: 978-1-59259-550-1

  • eBook Packages: Springer Protocols

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