Abstract
The antibody 2B4 combines with ferric mesoporphyrin to form an antibody-ferric mesoporphyrin complex which has a peroxidase activity. Formation of the complex was investigated by measuring the absorption in the Soret region after mixing the antibody and ferric mesoporphyrin. A rapid increase and a gradual decrease in the absorption were observed, and the respective first-order rate constants were obtained. From the dependence of values of the rate constants on the concentration of ferric mesoporphyrin, the complex formation was explained by a plausible mechanism, in which the antibody associated with ferric mesoporphyrin to form the first complex followed by a conformational change to the second complex. The first complex had almost the same peroxidase activity as that of the second complex. Our results suggests that the antibody acquires the peroxidase activity as soon as ferric mesoporphyrin is incorporated into its binding site, and that there will be no protein ligand to the iron center of ferric mesoporphyrin in the complex.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
REFERENCES
Abrams, J. J., and Webster, D. A. (1990). Arch. Biochem. Biophys. 279, 54–59.
Blackwood, M. E., Rush III, T. S., Romesberg, F., Schultz, P. G., and Spiro, T. G. (1998). Biochemistry 37, 779–782.
Campbell, D. A., Gong, B., Kochersperger, L. M., Yonkovich, S., Gallop, M. A., and Schultz, P. G. (1994). J. Am. Chem. Soc. 116, 2165–2166.
Choudhury, K., Sundaramoorthy, M., Hickman, A., Yonetani, T., Woehl, E., Dunn, M. F., and Poulos, T. L. (1994). J. Biol. Chem. 269, 20239–20249.
Cochran, A. G., and Schultz, P. G. (1990a). Science 249, 781–783.
Cochran, A. G., and Schultz, P. G. (1990b). J. Am. Chem. Soc. 112, 9414–9415.
Cowan, J. A. (1997). In Inorganic Biochemistry An Introduction, Second Edition, Wiley-VCH, New York, pp. 328–334.
Feng, Y., Liu, Z., Gao, G., Gao, S. J., Liu, X. Y., and Yang, T. S. (1995). Ann. N.Y. Acad. Sci. 750, 271–276.
Fersht, A. (1984). In Enzyme Structure and Mechanism, Second Edition, W. H. Freeman and Company, New York, pp. 128–143.
Foote, J., and Milstein, C. (1994). Proc. Natl. Acad. Sci. USA 91, 10370–10374.
Gramatikova, S. I., and Christen, P. (1996). J. Biol. Chem. 271, 30583–30586.
Hansson, U.-B., Wingren, C., and Alkner, U. (1997). Biochim. Biophys. Acta. 1340, 53–62.
Harada, A., Okamoto, K., Kamachi, M., Honda T., and Miwatani, T. (1990). Chem. Lett. 917–918.
Kawamura-Konishi, Y., Hosomi, N., Neya, S., Sugano, S., Funasaki, N., and Suzuki, H. (1996a). J. Biochem. 119, 857–862.
Kawamura-Konishi, Y., Neya, S., Funasaki, N., and Suzuki, H. (1996b). Biochem. Biophys. Res. Commun. 225, 537–544.
Kawamura-Konishi, Y., Asano, A., Yamazaki, M., Tashiro, H., and Suzuki, H. (1998). J. Mol. Catal. B: Enzymatic 4, 181–190.
Keinan, E., Sinha, S. C., Sinha-Bagchi, A., Benory, E., Ghozi, M. C., Eshhar, Z., and Green, B. S. (1990). Pure & Appl. Chem. 62, 2013–2019.
Kohda, K., Kakehi, M., Ohtsuji, Y., Tagaki, M., and Imanaka, T. (1997). FEBS Letters 407, 280–284.
Kranz, D. M., Herron, J. N., and Voss, Jr., E. W. (1982). J. Biol. Chem. 257, 6987–6995.
Lancet, D., and Pecht, I. (1976). Proc. Natl. Acad. Sci. USA 73, 3549–3553.
Pollack, S. J., Jacobs, J. W., and Schultz, P. G. (1986). Science 234, 1570–1573.
Quilez, R., de Lauzon, S., Desfosses, B., Mansuy, D., and Mahy, J.-P. (1996). FEBS Letters 395, 73–76.
Savitsky, A. P., Demcheva, M. V., Mantrova, E. Yu, and Ponomarev, G. V. (1994a). FEBS Letters 355, 314–316.
Savitsky, A. P., Nelen, M. I., Yatsmirsky, A. K., Demcheva, M. V., Ponomarev, G. V., and Sinikov, I. V. (1994b). Appl. Biochem. Biotechnol. 47, 317–327.
Schwabacher, A. W., Weinhouse, M. I., Auditor, M.-T. M., and Lerner, R. A. (1989). J. Am. Chem. Soc. 111, 2344–2346.
Shokat, K. M., Leumann, C. J., Sugasawara, R., and Schultz, P. G. (1988). Angew. Chem. Int. Ed. Engl. 27, 1172–1174.
Suzuki, H. (1994). J. Biochem. 115, 623–628.
Takagi, M., Kohda, K., Hamuro, T., Harada, A., Yamaguchi, H., Kamachi, M., and Imanaka, T. (1995). FEBS Letters 375, 273–276.
Tramontano, A., Janda, K. D., and Lerner, R. A. (1986). Science 234, 1566–1570.
Zidovetzki, R., Blatt, Y., Glaudemans, C. P. J., Manjula, B. N., and Pecht, I. (1980). Biochemistry 19, 2790–2795.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kawamura-Konishi, Y., Fujie, Y. & Suzuki, H. Kinetics of Formation of Antibody-Ferric Porphyrin Complex with Peroxidase Activity. J Protein Chem 18, 741–745 (1999). https://doi.org/10.1023/A:1020673316447
Published:
Issue Date:
DOI: https://doi.org/10.1023/A:1020673316447