The Anomeric Specificity of Yeast Pyruvate Kinase toward Activation by D-Fructose l,6-Bisphosphatete

Richard Fishbein, Patricia A. Benkovic, Stephen J. Benkovic

Research output: Contribution to journalArticle

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Abstract

The anomeric specificity of D-fructose 1,6-bisphosphate activation of yeast pyruvate kinase has been investigated utilizing stopped-flow kinetics and synthetic analogs of D-fructose 1,6-bisphosphate. It has been demonstrated that the immediate product, β-D-fructose 1,6-bisphosphate, of the phosphofructokinase-catalyzed reaction increases the catalytic activity of the enzyme. Although exclusive activation by the a anomer is ruled out by these experiments the possibility that the allosteric site is anomerically nonspecific cannot be excluded owing to experimental limits. 2,5-Anhydromannitol 1,6-bisphosphate, 2,5-anhydroglucitol 1,6-bisphosphate, 1,6-diphosphohexitol, and methyl (α+ β)-D-fructofuranoside 1,6-bisphosphate were tested as activators or inhibitors of the D-fructose 1,6-bisphosphate activation of pyruvate kinase. No activation was observed but inhibition of D-fructose 1,6-bisphosphate activation by 2,5-anhydromannitol 1,6-bisphosphate and 2,5-anhydroglucitol 1,6-bisphosphate was noted. Methyl (α + β)-D-fructofuranoside 1,6-bisphosphate and 1,6-diphosphohexitol also proved to inhibit weakly. The collective data suggest that the allosteric site on yeast pyruvate kinase may be nonspecific with respect to anomeric configuration, but that a C-2 hydroxyl is necessary for activation by D-fructose 1,6-bisphosphate.

Original languageEnglish (US)
Pages (from-to)4060-4063
Number of pages4
JournalBiochemistry
Volume14
Issue number18
DOIs
StatePublished - Sep 1 1975

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Pyruvate Kinase
Fructose
Yeast
Yeasts
Chemical activation
Allosteric Site
Phosphofructokinase-1
Phosphofructokinases
Hydroxyl Radical
fructose-1,6-diphosphate
Catalyst activity
Enzymes
Kinetics
Experiments

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Fishbein, Richard ; Benkovic, Patricia A. ; Benkovic, Stephen J. / The Anomeric Specificity of Yeast Pyruvate Kinase toward Activation by D-Fructose l,6-Bisphosphatete. In: Biochemistry. 1975 ; Vol. 14, No. 18. pp. 4060-4063.
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The Anomeric Specificity of Yeast Pyruvate Kinase toward Activation by D-Fructose l,6-Bisphosphatete. / Fishbein, Richard; Benkovic, Patricia A.; Benkovic, Stephen J.

In: Biochemistry, Vol. 14, No. 18, 01.09.1975, p. 4060-4063.

Research output: Contribution to journalArticle

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N2 - The anomeric specificity of D-fructose 1,6-bisphosphate activation of yeast pyruvate kinase has been investigated utilizing stopped-flow kinetics and synthetic analogs of D-fructose 1,6-bisphosphate. It has been demonstrated that the immediate product, β-D-fructose 1,6-bisphosphate, of the phosphofructokinase-catalyzed reaction increases the catalytic activity of the enzyme. Although exclusive activation by the a anomer is ruled out by these experiments the possibility that the allosteric site is anomerically nonspecific cannot be excluded owing to experimental limits. 2,5-Anhydromannitol 1,6-bisphosphate, 2,5-anhydroglucitol 1,6-bisphosphate, 1,6-diphosphohexitol, and methyl (α+ β)-D-fructofuranoside 1,6-bisphosphate were tested as activators or inhibitors of the D-fructose 1,6-bisphosphate activation of pyruvate kinase. No activation was observed but inhibition of D-fructose 1,6-bisphosphate activation by 2,5-anhydromannitol 1,6-bisphosphate and 2,5-anhydroglucitol 1,6-bisphosphate was noted. Methyl (α + β)-D-fructofuranoside 1,6-bisphosphate and 1,6-diphosphohexitol also proved to inhibit weakly. The collective data suggest that the allosteric site on yeast pyruvate kinase may be nonspecific with respect to anomeric configuration, but that a C-2 hydroxyl is necessary for activation by D-fructose 1,6-bisphosphate.

AB - The anomeric specificity of D-fructose 1,6-bisphosphate activation of yeast pyruvate kinase has been investigated utilizing stopped-flow kinetics and synthetic analogs of D-fructose 1,6-bisphosphate. It has been demonstrated that the immediate product, β-D-fructose 1,6-bisphosphate, of the phosphofructokinase-catalyzed reaction increases the catalytic activity of the enzyme. Although exclusive activation by the a anomer is ruled out by these experiments the possibility that the allosteric site is anomerically nonspecific cannot be excluded owing to experimental limits. 2,5-Anhydromannitol 1,6-bisphosphate, 2,5-anhydroglucitol 1,6-bisphosphate, 1,6-diphosphohexitol, and methyl (α+ β)-D-fructofuranoside 1,6-bisphosphate were tested as activators or inhibitors of the D-fructose 1,6-bisphosphate activation of pyruvate kinase. No activation was observed but inhibition of D-fructose 1,6-bisphosphate activation by 2,5-anhydromannitol 1,6-bisphosphate and 2,5-anhydroglucitol 1,6-bisphosphate was noted. Methyl (α + β)-D-fructofuranoside 1,6-bisphosphate and 1,6-diphosphohexitol also proved to inhibit weakly. The collective data suggest that the allosteric site on yeast pyruvate kinase may be nonspecific with respect to anomeric configuration, but that a C-2 hydroxyl is necessary for activation by D-fructose 1,6-bisphosphate.

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