Abstract
Calf intestinal alkaline phosphatase (CIP) was denatured in 3.0 M guanidine hydrochloride for 2 h at 25°C, before being diluted 20-fold with 0.1 M, pH 8.0, Tris-HCl buffer solution containing various effector molecules such as Mg2+, Zn2+, and nucleotide phosphate. The reactivation courses of the enzyme were investigated by the level of activity recovery, the recovery rate constant, and the relative standard deviation of the data. In the presence of effectors, the courses under reducing and nonreducing conditions of disulfide bonds of protein were compared. It was concluded that for CIP, Mg2+ is a more efficient inducer of reconstitution of the active site and appears to play a specific role. In addition, the present study discusses the differences in the refolding effectors between bacterial and mammalian enzymes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
REFERENCES
Coleman, J. E. (1992). Annu. Rev. Biophys. Biomol. Struct. 21: 441–453.
Ellis, R. J. (1991). Annu. Rev. Biochem. 60: 321–347.
Fernley, H. H. (1971). In: P. D. Boyer (Ed.), The Enzymes. 4, Academic Press, New York, pp. 417–447.
Fosset, M. D., and Chappelet-Tordo, M. L. (1974). Biochem. 13: 1783–1787.
Ghosh, N., Sarkar, S. N., and Roy, K. B. (1998). Biochem. 37: 15542–15547.
Kim, E. E., and Wyckoff, H. W. (1991). J. Mol. Biol. 281: 449–454.
Martin, J., Langer, T., and Boteva, R. (1991). Nature. 352: 36–42.
McComb, R. B., Bower, G. N., and Posen, S. (1979). Alkaline Phosphatase. Plenum Press, New York.
Murphy, J. E., and Kantrowitz, E. R. (1994). Mol. Microbiol. 12: 351–357.
Murphy, J. E., Xu, X., and Kantrowitz, E. R. (1993). J. Biol. Chem. 268: 21497–21500.
Rein, T. W., and Wilson, I. B. (1971). In: P. D. Boyer (Ed.), The Enzymes. 4, Academic Press, New York, pp. 374–415.
Sadis, S., Raghavendra, K., and Schuster, T. M. (1990). In Current Research in Protein Chemistry: Techniques, Structure and Function (Villafranca, J. J., Ed.), Academic Press, New York, pp. 339–347.
Sarkar, S. N., and Ghosh, N. (1996). Arch. Biochem. Biophys. 330: 174–180.
Subramaniam, V., Bergenhem, N. C., Gafni, A., and Steel, D. G. (1995). Biochem. 34: 1133–1136.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Tian, XJ., Song, XH., Yan, SL. et al. Study of Refolding of Calf Intestinal Alkaline Phosphatase. J Protein Chem 22, 417–422 (2003). https://doi.org/10.1023/B:JOPC.0000005456.69859.d9
Published:
Issue Date:
DOI: https://doi.org/10.1023/B:JOPC.0000005456.69859.d9