Abstract
Development of a reliable C-terminal sequencing method has been expected from various aspects including sequencing protein, analyzing posttranslational process, confirming recombinant proteins and cloning. Carboxypeptidase digestion has been commonly used with limitations. Since a classical isothiocyanate degradation was proposed, several modifications were reported in the past two conferences (Hawkes and Boyd, 1991; Inglis et al., 1991; Miller and Shively, 1989). The present paper summarizes two related novel carboxy-terminal sequencing methods using perfluorinated carboxylic acids in monohydrate form and carboxylic acid anhydrides. These reagents produce the mixtures of C-terminal successive degraded molecules which are able to be analyzed by fast-atom-bombardment (FAB)- or electrospray ionization (ESI)-mass spectrometry (MS).
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References
Hawke, D. H. and Boyd, V. L., 1991 Chemical C-terminal sequencing, in: Methods in Protein Sequence Analysis, Jörnvall, H., Höög, J.-O. and Gustavsson, A. M., eds., Birkhauser, Basel, 35.
Inglis, A. S., Moritz, R. L., Begg, G. S., Reid, G. E., Simpson, R. J. Graffunder, H., Matschull, L. and Wittmann-Liebold, B., 1991 C-terminal sequence analysis, in: Methods in Protein Sequence Analysis, Jörnvall, H., Höög, J.-O. and Gustavsson, A. M., eds., Birkhauser, Basel, 23.
Miller, C. G. and Shively, J. E., 1989 Carboxy-terminal sequence analysis of proteins and peptides by chemical methods, in: Methods in Protein Sequence Analysis, Wittmann-Liebold, B., ed., Springer-Verlag, Berlin, 145.
Tsugita, A., Takamoto, A., Kamo, M. and Iwadate, H., 1992a C-terminal sequencing of protein; a novel partial acid hydroloysis and analysis by mass spectrometry, Eur. J. Biochem. 206:691.
Tsugita, A., Takamoto, K. and Satake, K., 1992b Reaction of pentafluoropropionic anhydride vapor on polypeptide as revealed by mass spectrometry. A carboxypeptidase mimetic degradation, Chem. Lett. 235.
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© 1993 Springer Science+Business Media New York
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Tsugita, A. et al. (1993). Development of Novel C-Terminal Sequencing Methods. In: Imahori, K., Sakiyama, F. (eds) Methods in Protein Sequence Analysis. Springer, Boston, MA. https://doi.org/10.1007/978-1-4899-1603-7_7
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DOI: https://doi.org/10.1007/978-1-4899-1603-7_7
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