PMID: 7013791Mar 3, 1981Paper

Liberation of the triosephosphate isomerase reaction intermediate and its trapping by isomerase, yeast aldolase, and methylglyoxal synthase

Biochemistry
R Iyengar, I A Rose

Abstract

When a mixture of triosephosphate isomerase (rabbit muscle) and dihydroxyacetone phosphate (DHAP) is quenched with acid, a compound is liberated, presumed to be the cis-enediol 3-phosphate, that decomposes to inorganic phosphate (Pi) and methylglyoxal [Iyengar, R., & Rose, I.A. (1981) Biochemistry (preceding paper is this issue)]. The decomposition can be prevented by rapid neutralization if a catalytic amount of fresh isomerase is present. Varying the time between acidification and rescue gave a half-life of the liberate compound of approximately 12-17 ms. Varying the concentration of enzyme used for rescue gave a minimum second-order rate constant for trapping of 10(9)M(-1)s(-1). These results add further evidence favoring a stepwise mechanism for the aldose-ketose isomerase reactions in which a chemically defined enzyme-bound intermediate is found. The high rate of trapping over a wide pH range indicates that the enediol phosphate, not the enediolate phosphate, is the intermediate. One property of the enzyme is to stabilize the intermediate with respect to its fragmentation in solution by greater than 1000-fold. Yeast aldolase is also able to rescue all of the isomerase intermediate, though higher concentrations of enzyme ar...Continue Reading

References

Jun 1, 1976·Analytical Biochemistry·J P FroehlichR L Berger
Jun 22, 1977·Journal of the American Chemical Society·M C Summers, I A Rose
Jun 8, 1978·Nature·W G HolH J Berendsen
Dec 14, 1976·Biochemistry·W J Albery, J R Knowles
Aug 16, 1969·Nature·R Wolfenden
Feb 1, 1969·Biochemistry·R D KobesW J Rutter
Jun 1, 1969·Biochemistry·J F Riordan, P Christen
Jun 1, 1969·Biochemistry·C E HarrisW J Rutter

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Citations

Sep 1, 1983·Archives of Biochemistry and Biophysics·R M Mulligan, N E Tolbert
Feb 12, 2011·Seminars in Cell & Developmental Biology·Yoshiharu InoueWataru Nomura
Oct 26, 2000·The Journal of Biological Chemistry·E M FrickelB Mannervik
Jun 22, 2021·Annual Review of Biochemistry·Stephen J Benkovic

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