Triose phosphate isomerase reaction

The triose phosphate isomerase reaction completes the first phase of glycolysis, each glucose that passes through being converted to two molecules of glyceraldehyde-3-phosphate. Although the last two steps of the pathway are... 

The reaction mechanism is similar to the reaction promoted by phosphohexose isomerase in step (2) of glycolysis (Fig. 14-4). After the triose phosphate isomerase reaction, C-1, C-2, and C-3 of the starting glucose are chemically indistinguishable from C-6, C-5, and C-4, respectively (Fig. 14-6), setting up the efficient metabolism of the entire six-carbon glucose molecule. 

Fructose bisphosphate is cleaved by action of an aldolase (reaction 4) to give glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. These two triose phosphates are then equilibrated by triose phosphate isomerase (reaction 5 see also Chapter 13). As a result, both halves of the hexose can be metabolized further via glyceraldehyde 3-P to pyruvate. The oxidation of glyceraldehyde 3-P to the corresponding carboxylic acid, 3-phosphoglyceric acid (Fig. 17-7, reactions 6 and 7), is coupled to synthesis of a molecule of ATP from ADP and P . This means that two molecules of ATP are formed per hexose cleaved, and that two molecules of NAD+ are converted to NADH in the process. 

Answer Problem 1 outlines the steps in glycolysis involving fructose 1,6-bisphosphate, glyceraldehyde 3-phosphate, and dihydroxyacetone phosphate. Keep in mind that the aldolase reaction is readily reversible and the triose phosphate isomerase reaction catalyzes extremely rapid interconversion of its substrates. Thus, the label at C-l of glyceraldehyde 3-phosphate would equilibrate with C-l of dihydroxyacetone phosphate (AG ° = 7.5 kJ/mol). Because the aldolase reaction has AG ° = -23.8 kJ/mol in the direction of hexose formation, fructose 1,6-bisphosphate would be readily formed, and labeled in C-3 and C-4 (see Fig. 14-6). 

Figure 7.6. The metabolic reactions involved in the conversion of glycerol to glucose, the required precursor in the formation of sophorose. Note Reaction 1 catalyzed by triose phosphate isomerase. Reaction 2 catalyzed by aldolase. Reaction 3 catalyzed by fructose 1,6-bisphosphatase. Reaction 4 catalyzed by phosphoglucose isomerase., Reaction 6 catalyzed by glucose 6-phosphatase.

Triose Phosphate Isomerase (Reaction, Figure 1L9, Reaction, Reactions, Figure 11,10) Serine Protease (Table 5,4, Figure 11.11, Figure 1L12, Figure 11.13)... 

The reaction is similar to the hexose phosphate isomerase and triose phosphate isomerase reactions of glycolysis and probably proceeds through an enediol intermediate ... 

Stereospecifidly of Dihydroxyacetone Phosphate Reactions. In both the aldolase and triose phosphate isomerase reactions a carbon-hydrogen bond of the hydroxymethyl group is broken. Isotope exchange experiments with tritium have shown that both enzymes catalyze an equilibration between one hydrogen of the substrate and the hydrogen of water. ° The two enzymes do not attack the same hydrogen atom each is specific for only one position. In the projection shown (XII),... 

By measuring the relative intensities of the fructose-1,6-bisphosphate, dihydroxyacetone phosphate, and glyceraldehyde, it was possible to show that the aldolase reaction is in equilibrium in the cell. Similar measurements on dihydroxyacetone phosphate and glyceraldehyde-3-phosphate showed that the triose phosphate isomerase reaction is not in equilibrium. It was also found that the adenylate kinase reaction, 2ADP ATP + AMP, is in equilibrium in the intact cell without added oxygen or glucose. 

Further steps m glycolysis use the d glyceraldehyde 3 phosphate formed m the aldolase catalyzed cleavage reaction as a substrate Its coproduct dihydroxyacetone phosphate is not wasted however The enzyme triose phosphate isomerase converts dihydroxyacetone phosphate to d glyceraldehyde 3 phosphate which enters the glycol ysis pathway for further transformations... 

FIGURE 19.15 A reaction mechanism for triose phosphate isomerase. 

Triose phosphate isomerase catalyzes the conversion of dihy-droxyacetone-P to glyceraldehyde-3-P. The standard free energy change, AG°, for this reaction is +7.6 kj/mol. However, the observed free energy change (AG) for this reaction in erythrocytes is +2.4 kj/mol. 

The guilty party is the triose phosphate isomerase (TIM) reaction that interconverts DHAP and G3P. To be converted to pyruvate, the DHAP first has to be converted to G3P. TIM just moves the carbonyl group between the two carbons that don t have phosphate attached. TIM doesn t touch the phosphate. So, if the DHAP is labeled at the carbon that has the phosphate attached, the G3P that comes from DHAP will be labeled at the carbon with the phosphate attached. The carbon with the phosphate attached in the G3P that was produced directly by the aldolase reaction came from C-6 of glucose, but the carbon with the phosphate attached in the G3P that was produced from DHAP came from C-l of glucose. After TIM does it stuff, the carbon of G3P that has the phosphate will be... 

The natural substrate for the dehydrogenase, glyceraldehyde-3-phosphate (G-3-P), had been synthesized earlier by Hermann Fischer, Emil Fischer s son, and Baer in 1932. In 1934 Meyerhof and Lohmann synthesized hexose diphosphate, establishing it to be fructose 1,6 bisphosphate (F-l, 6 bis P). With F-1,6 bisP as substrate and hydrazine to trap the aldehydic and ketonic products of the reaction, G-3-P was identified in the mixture of G-3-P and dihydroxyacetone phosphate which resulted. Triose phosphate isomerase was then isolated and the importance of phosphorylated 3C derivatives established. 

We have seen many examples of chemical reactions involving enolate anions, and should now realize just how versatile they are in chemical synthesis (see Chapter 10). We have also seen several examples of how equivalent reactions are utilized in nature. For the triose phosphate isomerase mechanism above, we did not actually invoke a distinct enolate anion intermediate in the enolization process, but proposed that there was a smooth flow of electrons. For other reactions, we shall also need to consider whether enolate anions are actually involved, or whether a more favourable alternative exists. The aldol-type reaction... 

A quantitative expression developed by Albery and Knowles to describe the effectiveness of a catalyst in accelerating a chemical reaction. The function, which depends on magnitude of the rate constants describing individual steps in the reaction, reaches a limiting value of unity when the reaction rate is controlled by diffusion. For the interconversion of dihydroxacetone phosphate and glyceraldehyde 3-phosphate, the efficiency function equals 2.5 x 10 for a simple carboxylate catalyst in a nonenzymic process and 0.6 for the enzyme-catalyzed process. Albery and Knowles suggest that evolution has produced a nearly perfect catalyst in the form of triose-phosphate isomerase. See Reaction Coordinate Diagram... 

In what is now a classical study in enzyme kinetics, W. J. Albery and J. R Knowles developed a strategy for establishing a reaction coordinate diagram (shown in Fig. 2) for triose-phosphate isomerase catalysis using solvent exchange and kinetic isotope effect data. 

KINETIC ISOTOPE EEEECT REACTION COORDINATE DIAGRAM Triose-phosphate isomerase energetics, REACTION COORDINATE DIAGRAM TRIPLE-COMPETITIVE METHOD TRIPLET STATE FLUORESCENCE TRIPLET-TRIPLET ANNIHILATION ANNIHILATION... 

Interconversion between these three-carbon intermediates is a reversible reaction catalyzed by triose phosphate isomerase. 

As depicted in Figure 6.8 the stability screening was based on DERA activity assay, the retro-aldol reaction of 2-deoxy-D-ribose 5-phosphate to acetaldehyde and D-glyceraldehyde 3-phosphate. D-glyceraldehyde 3-phosphate is further converted by the auxiliary enzymes triose phosphate isomerase and glycerol phosphate dehydrogenase. As the latter reaction consumes NADH it can be measured spectro-pho to metrically by the decrease in absorbance at 340 nm. 

The 500-residue subunits of pyruvate kinase consist of four domains,891 the largest of which contains an 8-stranded barrel similar to that present in triose phosphate isomerase (Fig. 2-28). Although these two enzymes catalyze different types of reactions, a common feature is an enolic intermediate. One could imagine that pyruvate kinase protonates its substrate phosphoenolpyruvate (PEP) synchronously with the phospho group transfer (Eq. 12-42). However, the enzyme catalyzes the rapid conversion of the enolic form of pyruvate to the oxo form (Eq. 12-43) adding the proton sterospecifically to the si face. This and other evidence favors the enol as a true intermediate... 

Another detail should be mentioned. The active site of triose phosphate isomerase is formed by a series of loops connecting the a helices and (3 strands of the barrel. One of those loops, consisting of residues 167-176, folds over the active site after the substrate is bound to form a hinged lid that helps to hold the substrate in the correct orientation for reaction.150-152 When the lid, which can be seen in Fig. 13-6, closes, the peptide NH of G171 forms a hydrogen bond to a phosphate oxygen atom of the substrate. This is only one of many known enzymes with deeply buried active sites that close in some similar fashion before a rapid reaction occurs. 

Although enzymes tend to be extremely specific they are not always completely able to avoid side reactions. Triose phosphate isomerase releases small amounts of methylglyoxal (Eq. 13-28), presumably as... 

The rather toxic methylglyoxal is formed in many organisms and within human tissues.174 It arises in part as a side reaction of triose phosphate isomerase (Eq. 13-28) and also from oxidation of acetone (Eq. 17-7) or aminoacetone, a metabolite of threonine (Chapter 24).175 In addition, yeast and some bacteria, including E. coli, have a methylglyoxal synthase that converts dihydroxyacetone to methylglyoxal, apparently using a mechanism similar to that of triose phosphate isomerase. It presumably forms enediolate 2 of Eq. 13-26, which eliminates inorganic phosphate to yield methyl-... 

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