Abstract
Mouse brain proteins were separated by two-dimensional electrophoresis (2-DE). The proteins of a section of the 2-DE pattern were blotted onto hydrophobic membranes and 43 of them were excised and hydrolyzed by liquid-phase hydrolysis. The amino acid composition of these proteins was determined by orthophthaldialdehyde precolumn derivatization and compared with the compositions of known proteins stored in the NBRF sequence database. An identification program named ASA was developed for this purpose. The ASA program includes correction and weighting factors, data reduction by molecular weight windows, and exclusion or inclusion of certain organisms as desired. As a control, eight test proteins and five well-known proteins from mouse brain, all separated by 2-DE, were correctly identified by the program. Out of the 43 brain proteins selected, 19 were identified with high confidence.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aebersold, R., Teplow, D., Hood, L. E., and Kent, S. B. H. (1986).J. Biol. Chem. 261, 4229–4238.
Aebersold, R. H., Leavitt, J., Saavedra, R. A., Hood, L. E., and Kent, S. B. H. (1987).Proc. Natl. Acad. Sci. USA 84, 6970–6974.
Aebersold, R., and Leavitt, J. (1990).Electrophoresis 11, 517–527.
Anderson, N. L., Nance, S. L., Pearson, T. W., and Anderson, N. G. (1982).Electrophoresis 3, 135–142.
Ashman, K., and Bosserhoff, A. (1985). InModern Methods in Protein Chemistry—Review Articles (Tschesche, H., ed.), Walter de Gruyter, Berlin, Vol. 2, pp. 155–171.
Bauw, G., Van Damme, J., Puype, M., Vandekerckhove, J., Gesser, B., Ratz, G. P., Lauridsen, J. B., and Celis, J. E. (1989).Proc. Natl. Acad. Sci. USA 86, 7701–7705.
Chang, J. Y., Knecht, R., and Braun, D. G. (1983).Methods Enzymol. 91, 41–48.
Eckerskorn, C., Jungblut, P., Mewes, W., Klose, J., and Lottspeich, F. (1988a).Electrophoresis 9, 830–838.
Eckerskorn, C., Mewes, W., Goretzki, H., and Lottspeich, F. (1988b).Eur. J. Biochem. 176, 509–519.
Eckerskorn, C., and Lottspeich, F. (1989).Chromatographia 28, 92–94.
Einarsson, S., Josefsson, B., and Lagerkvist, S. (1983).J. Chromatogr. 282, 609–618.
Hirano, H., and Watanabe, T. (1990).Electrophoresis 1, 573–580.
Jungblut, P., Choli, T., and Wittmann-Liebold, B. (1989).Biol. Chem. Hoppe-Seyler 370, 775.
Jungblut, P., Eckerskorn, C., Lottspeich, F., and Klose, J. (1990).Electrophoresis 11, 581–588.
Jungblut, P.et al., in preparation (1992).
Kennedy, T. E., Gawinowicz, M. A., Barzilai, A., Kandel, E. R., and Sweat, J. D. (1988).Proc. Natl. Acad. Sci. USA 85, 7008–7012.
Klose, J. (1975).Humangenetik 26, 231–243.
Klose, J. (1983). InModern Methods in Protein Chemistry—Review Articles (Tschesche, H., ed.), Walter de Gruyter, Berlin, pp. 49–78.
Laemmli, U. K. (1979).Nature 227, 680–685.
Matsudaira, P. (1987).J. Biol. Chem. 262, 10,035–10,038.
Ozols, J. (1990).Methods in Enzymol. 182, 587–601.
Peterson, G. L. (1977).Anal. Biochem. 83, 346–356.
Sibbald, P. R., Sommerfeldt, H., and Argos, P. (1991).Anal. Biochem. 198, 330–333.
Tous, G. I., Fausnaugh, J. L., Akinyosoye, O., Lackland, H., Winter-Cash, P., Vitorica, F. J., and Stein, S. (1989).Anal. Biochem. 179, 50–55.
Turnell, D. C., and Cooper, J. D. H. (1982).Clin. Chem. 28, 527–531.
Ui, N. (1971).Biochim. Biophys. Acta 229, 567–581.
Vandekerckhove, J., Bauw, G., Puype, M., Van Damme, J., and Van Montagu, M. (1985).Eur. J. Biochem. 152, 9–19.
Walsh, M., McDougall, J., and Wittmann-Liebold, B. (1988).Biochemistry 27, 6867–6876.
Watanabe, Y., and Imai, K. (1981).Anal. Biochem. 136, 471–474.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Jungblut, P., Dzionara, M., Klose, J. et al. Identification of tissue proteins by amino acid analysis after purification by two-dimensional electrophoresis. J Protein Chem 11, 603–612 (1992). https://doi.org/10.1007/BF01024960
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01024960