Abstract
Anaerobiospirillum succiniciproducens His225Gln, Asp262Asn, Asp263Asn, and Thr249Asn phosphoenolpyruvate carboxykinases were analyzed for their oxaloacetate decarboxylase, and pyruvate kinase–like activities. The His225Gln and Asp263Asn enzymes showed increased K m values for Mn2+ and PEP compared with the native enzyme, suggesting a role of His225 and Asp263 in Mn2+ and PEP binding. No mayor alterations in K m values for oxaloacetate were detected for the varied enzymes. Alterations of His225, Asp262, Asp263, or Thr249, however, did not affect the V max of the secondary activities as much as they affected the V max for the main reaction. The results presented in this communication suggest different rate-limiting steps for the primary reaction and the secondary activities.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
REFERENCES
Ash, D. E., Emig, F. A., Chowdhury, S. A., Satoh, Y., and Schramm, V. L. (1990). J. Biol. Chem. 265: 7377-7384.
Jabalquinto, A. M., Laivenieks, M., Zeikus, J. G., and Cardemil, E. (1999). J. Protein Chem. 18: 659-664.
Jabalquinto, A. M., Laivenieks, M., González-Nilo, F. D., Yevenes, A., Encinas, M. V., Zeikus, J. G., et al. (2002). J. Protein Chem. 21: 393-401.
Krautwurst, H., Bazaes, S., González, F. D., Jabalquinto, A. M., Frey, P. A., and Cardemil, E. (1998). Biochemistry 37: 6295-6302.
Llanos, L., Briones, R., Yévenes, A., González-Nilo, F. D., Frey, P. A., and Cardemil, E. (2001). FEBS Lett. 493: 1-5.
Martel, A. E., and Smith, R. M. (1998). NIST Critically Selected Stability Constants of Metal Complexes, NIST standard references database 46 version 5.0.
Matte, A., Tari, L. W., Goldie, H., and Delbaere, L. T. J. (1997). J. Biol. Chem. 272: 8105-8108.
Noce, P. S., and Utter, M. F. (1975). J. Biol. Chem. 250: 9099-9105.
Parker, D. R., Norvell, W. A., and Chaney, R. L. (1995). GEOCHEMPC: A chemical speciation program for IBM and compatible personal computers. In: R. H. Loeppert, et al. (eds.), Chemical Equilibrium and Reaction Models, SSSA Spec. Publ. No. 42, American Society of Agronomy, Madison, WI.
Tari, L. W., Matte, A., Goldie, H., and Delbaere, L. T. J. (1997). Nat. Struct. Biol. 4: 990-994.
Utter, M. F., and Kolenbrander, H. M. (1972). The Enzymes (3rd Ed., Vol. 6), Academic Press, New York, pp. 117-168.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Jabalquinto, A.M., Laivenieks, M., Cabezas, M. et al. The Effect of Active Site Mutations in the Oxaloacetate Decarboxylase and Pyruvate Kinase-Like Activities of Anaerobiospirillum succiniciproducens Phosphoenolpyruvate Carboxykinase. J Protein Chem 21, 443–445 (2002). https://doi.org/10.1023/A:1021342918955
Published:
Issue Date:
DOI: https://doi.org/10.1023/A:1021342918955