Abstract
A new procedure to selectively identify disulfide-containing peptides in extracts of biological tissues is described. Disulfide-containing peptides are detected by their UV absorbance and electrochemical (EC) activity after chromatographic separation, and subsequently identified by fast atom bombardment mass spectrometry (FABMS). This combination of fractionation by HPLC and selective detection is attractive because it is rapid, highly specific for disulfide-containing peptides, and applicable to all disulfide-containing peptides that may be present in complex biological mixtures. Useful procedures for applying the method are demonstrated with tissue extracts from bovine pituitary and catfish pancreas. In addition to finding the expected disulfide-containing peptides, evidence for two forms of catfish insulin are presented. The merits of this and other methods used to detect peptides in similar tissue extracts are discussed.
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References
Allison, L. A., and Shoup, R. E. (1983).Anal. Chem. 55, 8–12.
Andrews, P. C. (1989). InCurrent Research in Protein Chemistry (Villafranca, J., ed.), Academic Press, New York (in press).
Andrews, P. C., and Dixon, J. E. (1981).J. Biol. Chem. 256, 8267–8270.
Andrews, P. C., and Ronner, P. (1985).J. Biol. Chem. 260, 3910–3914.
Andrews, P. C., Pubols, M. H., Hermodson, M. A., Sheares, B. T., and Dixon, J. E. (1984).J. Biol. Chem. 259, 13267–13272.
Andrews, P. C., Mershid, A., and Cotter, R. J. (1988).Anal. Biochem. 174, 23–31.
Creighton, T. E. (1984). InMethods in Enzymology, Vol. 107 (Wold, F., and Moldave, K., eds.), Academic Press, New York, pp. 305–329.
Feistner, G. J., Hojrup, P., Evans, C. J., Barofsky, D. F., Faull, K. F., and Roepstorff, P. (1989).Proc. Natl. Acad. Sci. USA 86, 6013–6017.
Garvie, C. T., Straub, K. M., and Lynn, R. K. (1987).J. Chromatogr. 413, 43–52.
Haeffrer-Gormley, L., Parente, L., and Wetlaufer, D. B. (1985).Int. J. Pept. Protein Res. 26, 83–91.
Lazure, C., Rochemont, J., Seidah, N. G., and Chretien, M. (1985).J. Chromatogr. 326, 339–348.
Morris, H. R., and Pucci, P. (1985).Biochem. Biophys. Res. Commun. 126, 1122–1128.
Nakanishi, F., Inoue, A., Kito, T., Nakamura, M., Chang, A. C. Y., Cohen, F. N., and Numa, F. (1979).Nature 278, 423–427.
Oyama, H., Bradshaw, R. A., Bates, O. J., and Permutt, A. (1980).J. Biol. Chem. 255, 2251–2254.
Pettinga, C. W. (1958). InBiochemical Preparations, Vol. 6 (Vestling, C. S., ed.), Wiley, New York, pp. 28–31.
Richie J. P., Jr., and Lang, C. A. (1987).Anal. Biochem. 163, 9–15.
Sun, Y., and Smith, D. L. (1988).Anal. Biochem. 172, 130–138.
Yazdanparast, R., Andrews, P. C., Smith, D. L., and Dixon, J. E., (1987).J. Biol. Chem. 262, 2507–2513.
Yazdanparast, R., Andrews, P. C., Smith, D. L., and Dixon, J. E. (1986).Anal. Biochem. 153, 348–353.
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Sun, Y., Andrews, P.C. & Smith, D.L. Identification of disulfide-containing peptides in endocrine tissue extracts by HPLC-electrochemical detection and mass spectrometry. J Protein Chem 9, 151–157 (1990). https://doi.org/10.1007/BF01025306
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DOI: https://doi.org/10.1007/BF01025306