Abstract
The rate of oxidation of ferricyanide of the aldolase-dihydroxyacetone phosphate complex was measured under different conditions. The following conclusions are drawn. 1. In the cleavage of fructose diphosphate, catalysed by native aldolase, the steady-state concentration of the enzyme-dihydroxyacetone phosphate carbanion intermediate represents less than 6% of the total enzyme-substrate intermediates. 2. Fructose diphosphate and dihydroxyacetone phosphate compete for the four catalytic sites on aldolase, the binding of fructose diphosphate being about twice as tight. 3. The equilibrium concentration of the carbanion intermediate formed by reaction of carboxypeptidase-treated aldolase with dihydroxyacetone phosphate is independent of pH between 5.0 and 9.0. The rates of fromation of the carbanion intermediate and of the reverse reaction are, however, concomitantly increased by increasing pH between 5.0 and 6.5.
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- Christen P., Riordan J. F. Reaction of an aldolase-substrate intermediate with tetranitromethane. Biochemistry. 1968 Apr;7(4):1531–1538. doi: 10.1021/bi00844a041. [DOI] [PubMed] [Google Scholar]
- Ginsburg A., Mehler A. H. Specific anion binding to fructose diphosphate aldolase from rabbit muscle. Biochemistry. 1966 Aug;5(8):2623–2634. doi: 10.1021/bi00872a021. [DOI] [PubMed] [Google Scholar]
- Grazi E. On the substrate specificity of fructose 1,6-diphosphate aldolase (E.C.4.1.2.13) from rabbit muscle: a critical revision. Biochem Biophys Res Commun. 1974 Jul 10;59(1):450–454. doi: 10.1016/s0006-291x(74)80227-5. [DOI] [PubMed] [Google Scholar]
- Grazi E. Quantitative evaluation of the carbanion intermediate in the aldolase reaction. Biochem Biophys Res Commun. 1974 Jan;56(1):106–111. doi: 10.1016/s0006-291x(74)80321-9. [DOI] [PubMed] [Google Scholar]
- Grazi E., Sivieri-Pecorari C., Gagliano R., Trombetta G. Complexes of fructose diphosphate aldolase with dihydroxyacetone phosphate and dihydroxyacetone sulfate. Biochemistry. 1973 Jul 3;12(14):2583–2590. doi: 10.1021/bi00738a006. [DOI] [PubMed] [Google Scholar]
- Grazi E., Trombetta G. Fructose-bisphosphate aldolase from rabbit muscle. A thermodynamic study on the formation of the enzyme-dihydroxyacetone phosphate complex. Biochim Biophys Acta. 1974 Sep 11;364(1):120–127. doi: 10.1016/0005-2744(74)90139-9. [DOI] [PubMed] [Google Scholar]
- HORECKER B. L., ROWLEY P. T., GRAZI E., CHENG T., TCHOLA O. THE MECHANISM OF ACTION OF ALDOLASES. IV. LYSINE AS THE SUBSTRATE-BINDING SITE. Biochem Z. 1963;338:36–51. [PubMed] [Google Scholar]
- Healy M. J., Christen P. Mechanistic probes for enzymatic reactions. Oxidation-reduction indicators as oxidants of intermediary carbanions (studies with aldolase, aspartate aminotransferase, pyruvate decarboxylase, and 6-phosphogluconate dehydrogenase). Biochemistry. 1973 Jan 2;12(1):35–41. doi: 10.1021/bi00725a006. [DOI] [PubMed] [Google Scholar]
- Kawahara K., Tanford C. The number of polypeptide chains in rabbit muscle aldolase. Biochemistry. 1966 May;5(5):1578–1584. doi: 10.1021/bi00869a018. [DOI] [PubMed] [Google Scholar]
- ROSE I. A., O'CONNELL E. L., MEHLER A. H. MECHANISM OF THE ALDOLASE REACTION. J Biol Chem. 1965 Apr;240:1758–1765. [PubMed] [Google Scholar]
- RUTTER W. J., RICHARDS O. C., WOODFIN B. M. Comparative studies of liver and muscle aldolase. I. Effect of carboxypeptidase on catalytic activity. J Biol Chem. 1961 Dec;236:3193–3197. [PubMed] [Google Scholar]
- SPOLTER P. D., ADELMAN R. C., WEINHOUSE S. DISTINCTIVE PROPERTIES OF NATIVE AND CARBOXYPEPTIDASE-TREATED ALDOLASES OF RABBIT MUSCLE AND LIVER. J Biol Chem. 1965 Mar;240:1327–1337. [PubMed] [Google Scholar]