Abstract
A kininogenase from bovine erythrocyte membranes has been purified 140-fold by affinity chromatography on pepstatin A-Agarose followed by ion exchange chromatography on CM Cellulose. The purified enzyme showed an apparent molecular weight of 31,000 daltons as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. ItspH optimum is 7.5, and it was totally inhibited by soybean trypsin inhibitor, phenylmethylsulfonylfluoride, aprotinin, pepstatin, and dithiotreitol, suggesting the presence of a disulfide bond(s) whose integrity is(are) essential for maintaining the native three-dimensional structure. The referred enzyme was able to release kinin from a substrate partially purified from rat plasma. The kininogenase was activated by Zn2+, Ca2+, and cysteine-HCl.
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References
Barlas, A., Sugio, K., and Greenbaum, L. M. (1985) Release of T-kinin and bradykinin in carrageenin induced inflammation in the rat,FEBS Lett. 190, 268–270.
Damas, J., Remacle-Volon, G., and Deflandre, E. (1986). Further studies of the mechanism of counter irritation by turpentine.Naunyn-Schmiedeberg's Arch. Pharmacol. 332, 196–200.
Friberger, P., Aurell, L., and Claeson, G. (1982). Chromogenic substrate of kallikrein and related enzymes,Agents Actions 9, 83–90.
Gutman, N., Moreau, T., Alhenc-Gelas, F., Baussant, T., Moujahed, A., Akpona, S., and Gauthier, F. (1988). T-kinin release from T-kininogen by rat-submaxillary-gland endopeptidase K,Eur. J. Biochem. 171, 577–582.
Jupp, R. A., Richards, A. D., Kay, J., Dunn, B. M., Wyckoff, J. B., Samloff, M. I., and Yamamoto, K. (1988). Identification of the aspartic proteinases from human erythrocyte membranes and gastric mucosa (slow-moving proteinase) as catalytically equivalent to cathepsin E,Biochem. J. 254, 895–898.
Laemmli, U. K. (1970). Cleavage structural proteins during the assembly of the head of bacteriophage T4,Nature 217, 680–685.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951). Protein measurement with the Folin phenol reagent,J. Biol. Chem. 193, 265–275.
Morrison, W. L., and Neurath, H. (1953). Proteolytic enzymes of the formed elements of human blood,J. Biol. Chem. 200, 39–51.
Ryle, A. P. (1970). The porcine pepsins and pepsinogens. InMethods in Enzumology, Vol. XIX (Perlmann, G. E., and Lorand, L., eds.), Academic Press, New York, pp. 316–346.
Yamamoto, K., and Marchesi, V. (1984). Purification and characterization of acid proteinase from human erythrocyte membranes,Biochem. Biophys. Acta 790, 208–218.
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Ferreira, L.A.F., Bergamasco, M. & Henriques, O.B. Isolation and properties of a T-kininogenase from bovine erythrocyte membranes. J Protein Chem 13, 547–552 (1994). https://doi.org/10.1007/BF01901536
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DOI: https://doi.org/10.1007/BF01901536