Abstract
The influence of transketolase substrates on the interaction of apotransketolase with its coenzyme thiamine diphosphate (TDP) and on the stability of the reconstituted holoenzyme was studied. Donor substrates increased the affinity of the coenzyme for transketolase, whereas acceptor substrate did not. In the presence of magnesium ions, the active centers of transketolase initially identical in TDP binding lose their equivalence in the presence of donor substrates. The stability of transketolase depended on the cation type used during its reconstitution—the holoenzyme reconstituted in the presence of calcium ions was more stable than the holoenzyme produced in the presence of magnesium ions. In the presence of donor substrate, the holoenzyme stability increased without depending on the cation used during the reconstitution. Donor substrate did not influence the interaction of apotransketolase with the inactive analog of the coenzyme N3′-pyridyl thiamine diphosphate and did not stabilize the transketolase complex with this analog. The findings suggest that the effect of the substrate on the interaction of the coenzyme with apotransketolase and on stability of the reconstituted holoenzyme is caused by generation of 2-(α,β-dihydroxyethyl)thiamine diphosphate (an intermediate product of the transketolase reaction), which has higher affinity for apotransketolase than TDP.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- TK:
-
transketolase
- TDP:
-
thiamine diphosphate
- DHETDP:
-
2-(α,β-dihydroxyethyl)thiamine diphosphate
REFERENCES
Kochetov, G. A. (1982) Meth. Enzymol., 90, 209–223.
Kochetov, G. A., and Philippov, P. P. (1970) Biochem. Biophys. Res. Commun., 38, 930–933.
Nikkola, M., Lindqvist, Y., and Schneider, G. (1994) J. Biol. Chem., 238, 387–404.
Sundstrom, M., Lindqvist, Y., and Schneider, G. (1992) FEBS Lett., 313, 229–231.
Kochetov, G. A., Tikhomirova, N. K., and Philippov, P. P. (1975) Biochem. Biophys. Res. Commun., 63, 924–930.
Egan, R. M., and Sable, H. Z. (1981) J. Biol. Chem., 256, 4877–4883.
Kovina, M. V., Selivanov, V. A., Kochevova, N. V., and Kochetov, G. A. (1998) Biochemistry (Moscow), 63, 988–995.
Kovina, M. V., Selivanov, V. A., Kochevova, N. V., and Kochetov, G. A. (1997) FEBS Lett., 418, 11–14.
Selivanov, V. A., Kovina, M. V., Kochevova, N. V., Meshalkina, L. E., and Kochetov, G. A. (2003) J. Mol. Catal. B: Enzymatic, 26, 33–40.
Heinrich, P. C., Steffen, H., Janser, P., and Wiss, O. (1972) Eur. J. Biochem., 30, 533–541.
Heinrich, P. C., and Schmidt, D. (1973) Experientia, 29, 1226–1227.
Lindqvist, Y., Schneider G., Ermler, U., and Sundstrom, M. (1992) EMBO J., 11, 218–223.
Kochetov, G. A., and Izotova, A. E. (1973) Biokhimiya, 38, 954–957.
Fiedler, E., Thorell, S., Sandalova, T., Golbik, R., Konig, S., and Schneider, G. (2002) Proc. Natl. Acad. Sci. USA, 99, 591–595.
Schellenberger, A., Wendler, K., Creutzburg, P., and Hubner, G. (1967) Hoppe-Seyler’s Z. Physiol. Chem., 348, 501–508.
Wikner, C., Meshalkina, L., Nilsson, U., Nikkola, M., Lindqvist, Y., Sundstrom, M., and Schneider, G. (1994) J. Biol. Chem., 269, 32144–32150.
Heinrich, C. P., Noack, K., and Wiss, O. (1972) Biochem. Biophys. Res. Commun., 49, 1427–1432.
Kochetov, G. A., Usmanov, R. A., and Mevkh, A. T. (1973) Biochem. Biophys. Res. Commun., 54, 1619–1626.
Kochetov, G. A., Meshalkina, L. E., and Usmanov, R. A. (1976) Biochem. Biophys. Res. Commun., 69, 836–843.
Meshalkina, L. E., and Kochetov, G. A. (1979) Dokl. Akad. Nauk SSSR, 248, 1482–1486.
Meshalkina, L. E., Usmanov, R. A., and Kochetov, G. A. (1990) in Biochemistry and Physiology of Thiamine Diphosphate Enzymes (Bisswanger, H., and Ullrich, J., eds.) Verlag Chemie, Weinheim, pp. 343–352.
Dixon, M. (1972) Biochem. J., 129, 197–203.
Usmanov, R., Sidorova, N., and Kochetov, G. (1996) Biochem. Mol. Biol. Int., 38, 307–314.
Pustynnikov, M. G., Neef, H., Usmanov, R. A., Shellenberger, A., and Kochetov, G. A. (1986) Biokhimiya, 51, 1003–1016.
Meshalkina, L. E., Golbik, R., Usmanov, R. A., Neef, H., Shellenberger, A., and Kochetov, G. A. (1989) Dokl. Akad. Nauk SSSR, 307, 486–490.
Konig, S., Schellenberger, A., Neef, H., and Schneider, G. (1994) J. Biol. Chem., 269, 10879–10882.
Hennig, J., Kern, G., Neef, H., Bisswanger, H., and Hubner, G. (1996) in Biochemistry and Physiology of Thiamine Diphosphate Enzymes (Bisswanger, H., and Ullrich, J., eds.) Verlag Chemie, Weinheim, pp. 235–242.
Hubner, G., Kern, G., Hennig, J., Neef, H., and Bisswanger, H. (1996) in Biochemistry and Physiology of Thiamine Diphosphate Enzymes (Bisswanger, H., and Ullrich, J., eds.) Verlag Chemie, Weinheim, pp. 243–251.
Yi, J., Nemeria, N., McNally, A., and Jordan, F. (1996) J. Biol. Chem., 271, 33192–33200.
Sumegi, B., and Alkonyi, I. (1983) Eur. J. Biochem., 136, 347–353.
Author information
Authors and Affiliations
Corresponding author
Additional information
__________
Translated from Biokhimiya, Vol. 70, No. 7, 2005, pp. 933–940.
Original Russian Text Copyright © 2005 by Esakova, Khanova, Meshalkina, Golbik, Hubner, Kochetov.
Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM04-131, December 5, 2004.
Rights and permissions
About this article
Cite this article
Esakova, O.A., Khanova, E.A., Meshalkina, L.E. et al. Effect of Transketolase Substrates on Holoenzyme Reconstitution and Stability. Biochemistry (Moscow) 70, 770–776 (2005). https://doi.org/10.1007/s10541-005-0182-4
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/s10541-005-0182-4