Abstract
Although a variety of methods have been available for coupling proteins with water-insoluble carriers, the impetus to our experiments was given by a publication of Levin et al. (1). They coupled trypsin to the copolymer of maleic anhydride and ethylene (EMA) using hexamethylenediamine as cross-linking agent. The proposed mechanism of action involves the binding of the enzyme covalently via the ε-amino group of lysine in the protein. (Presumably the other protein binding agents used in our studies also acted this way or by linking tyrosines of proteins.) Levin’s method was successfully applied by Fritz et al. (2, 3), who used it for purification of enzymes and inhibitors. By coupling plasmin or trypsin to EMA and subsequently reacting them with inhibitors present in crude extracts, they separated and purified the inhibitors. The purification of kallikrein was achieved by adsorbing the enzyme on an EMA-Trasylol column and subsequently eluting it with guanidine solution.
These studies were supported in part by a grant (HE 08764) from National Institutes of Health, U. S. P. H. S. and by the O. N. R. (N00014-68-A-0496).
During the tenure of the fellowship of the Oklahoma Heart Association
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Levin, Y., M. Pecht, L. Goldstein and E. Katchalski, Biochem. 3: 1905, 1964.
Fritz, H., M. Gebhardt, E. Fink, W. Schramm and E. Werle, Hoppe-Seyler’s Z. Physiol. Chem. 350: 129, 1969.
Fritz, H., B. Brey, A. Schmal and E. Werle, Hoppe-Seyler’s Z. Physiol. Chem. 350: 617, 1969.
Chiang, T. S., E. G. Erdb’s, I. Miwa, L. L. Tague and J. J. Coalson. Circ. Res. 23: 507, 1968.
Erdös, E G., T. Seki, H. Y. T. Yang and L. L. Tague. Pharmacol. Res. Commun. 1: 152, 1969.
Axen, R., J. Porath and S. Ernback. Nature 214: 1302, 1967.
Cuatrecasas, P., M. Wilchek and C. B. Anfinsen. Proc. Nat. Acad. Sci. 61: 636, 1968.
Haynes, R. and K. A. Walsh. Biochem. Biophys. Res. Commun. 36: 235, 1969.
Erdb’s, E. G., L. L. Tague and I. Miwa. Biochem. Pharmacol. 17: 667, 1968.
Werle, E. in “Polypeptides which affect smooth muscles and blood vessels”, M. Schacter, ed., p. 199. Pergamon Press, Oxford, 1960.
Bar-Eli, A. and E. Katchalski. J. Biol. Chem. 238: 1690, 1963.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1970 Plenum Press, New York
About this chapter
Cite this chapter
Seki, T., Yang, H.Y.T., Levin, Y., Jenssen, T.A., Erdös, E.G. (1970). Application of Water-Insoluble Complexes of Kininogenases, Inhibitors and Kininases to Kinin Research. In: Sicuteri, F., e Silva, M.R., Back, N. (eds) Bradykinin and Related Kinins. Advances in Experimental Medicine and Biology, vol 8. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-3198-8_4
Download citation
DOI: https://doi.org/10.1007/978-1-4684-3198-8_4
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4684-3200-8
Online ISBN: 978-1-4684-3198-8
eBook Packages: Springer Book Archive