Abstract
Peptides, cleaved by a mixture of carboxypeptidases CPP and CPY, can be detected by MALDI MS and the amino acid sequence thereby determined by calculation of the differences between consecutive peaks. In the present study we have used derivatizations of Lys and Cys to facilitate identification of these residues. Since the mass values do not readily distinguish Lys from Gln, we have converted Lys to homoarginine by guanidination, allowing simple detection of Lys. To identify the Cys positions in peptides that contain cystine, cysteic acid, or carboxymethylcysteine is not possible using CPY and CPP because of the lack of proteolytic cleavage. Instead we find that identification of Cys residues within the sequence can be achieved after conversion to a basic derivative, 4-thialaminine (Thi), by trimethylaminoethylation.
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REFERENCES
Bradley, C. V., Williams, D. H., and Hanley, M. R. (1982). Biochem. Biophys. Res. Commun. 104, 1223–1230.
Breddam, K. (1986). Carlsberg Res. Commun. 51, 83–128.
Chait, B. T., Chaudhary, T., and Field, F. H. (1987). In Methods in Protein Sequence Analysis (Walsh, K. A., eds.), Humana Press, Clifton, New Jersey, pp. 483–492.
Glazer, A. N., DeLange, R. J., and Sigman, D. S. (1975). In Laboratory Techniques in Biochemistry and Molecular Biology (Work, T. S., and Work, E., eds), Elsevier, Amsterdam.
Itano, H. A., and Robinson, E. A. (1972). J. Biol. Chem. 247, 4819–4824.
Kallweit, U., Börnsen, K. O., Kresbach, G. M., and Widmer, H. M. (1996). Rapid Commun. Mass Spectrom. 10, 845–849.
Kimmel, J. R. (1967). Meth. Enzymol. 11, 584–589.
Liepinsh, E., Berndt, K. D., Sillard, R., Mutt, V., and Otting, G. (1994). J. Mol. Biol. 239, 137–153.
Mutt, V. (1959). Arkiv Kemi 15, 69–74.
Mutt, V. (1978). In Gut Hormones (Bloom, S. R., ed.), Churchill Livingstone, Edinburgh.
Nguyen, D. N., Becker, G. W., and Riggin, R. M. (1995). J. Chromatogr. 705, 21–45.
Rosnack, K. J., and Stroh, J. G. (1992). Rapid Commun. Mass Spectrom. 6, 637–640.
Schär, M., Börnsen, K. O., and Gassmann, E. (1991). Rapid Commun. Mass Spectrom. 5, 319–326.
Stults, J. T. (1995). Curr. Opin. Struct. Biol. 5, 691–698.
Thiede, B., Wittmann-Liebold, B., Bienert, M., and Krause, E. (1995). FEBS Lett. 357, 65–69.
Woods, A. S., Huang, A. Y., Cotter, R. J., Pasternack, G. R., Pardoll, D. M., and Jaffee, E. M. (1995). Anal. Biochem. 226, 15–25.
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Bonetto, V., Bergman, AC., Jörnvall, H. et al. C-Terminal Sequence Determination of Modified Peptides by MALDI MS. J Protein Chem 16, 371–374 (1997). https://doi.org/10.1023/A:1026328520307
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DOI: https://doi.org/10.1023/A:1026328520307