Interactions of fructose 1,6-diphosphate, substrates, and monovalent cations with yeast pyruvate kinase monitored by changes in enzyme fluorescence

Kuczenski, Ronald; Suelter, Clarence H.

doi:10.1021/bi00791a010

Cited by 25 publications

(16 citation statements)

References 23 publications

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“…In experiments where the effector concentrations were generally less than 1 mm, they showed that the dissociation constant of fructose bisphosphate was decreased 2.5-fold when 5 mM-ADP was added to solutions of the enzyme that contained 25 mM-Mg2+. Kuczenski & Suelter (1971a) regarded this result (which is in agreement with our findings) as surprising because of the observed insensitivity of ADP binding to an addition of 1 mM-fructose bisphosphate (Hunsley & Suelter 1969), which we referred to above when discussing Fit D1200. This is, therefore, an independent recognition of the possibility that the influence of fructose bisphosphate on yeast pyruvate kinase is fundamentally different at high and at low concentrations.…”

Section: Modelling the Datasupporting

confidence: 88%

“…The independence of the maximum velocity from the fructose bisphosphate concentration is already well-established for the yeast enzyme (Haeckel et al, 1968;Hunsley & Suelter, 1969;Johannes & Hess, 1973) and for the enzymes from rabbit muscle (Phillips & Ainsworth, 1977) and liver (Irving & Williams, 1973). The observation permits the conclusion that addition of effector does not decrease the molecular mass of the enzyme with its consequent inactivation (Kuczenski & Suelter, 1970, 1971a.…”

Section: Modelling the Datasupporting

confidence: 56%

“…The positive values of kAD, kBD and kCD also require the complementary result, that is that the affinity of the enzyme for fructose bisphosphate should be increased by saturation with the substrates. Kuczenski & Suelter (1971a) have demonstrated this effect in fluorimetric studies of fructose bisphosphate binding by yeast pyruvate kinase at pH 7.5. In experiments where the effector concentrations were generally less than 1 mm, they showed that the dissociation constant of fructose bisphosphate was decreased 2.5-fold when 5 mM-ADP was added to solutions of the enzyme that contained 25 mM-Mg2+.…”

Section: Modelling the Datamentioning

confidence: 85%

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The regulatory properties of yeast pyruvate kinase. Effect of fructose 1,6-bisphosphate

Morris¹,

Ainsworth²,

Kinderlerer³

1986

Biochemical Journal

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The kinetics of pyruvate kinase from Saccharomyces cerevisiae were studied in assays at pH 6.2 at 25 degrees C as a function of the concentrations of the substrates ADP, phosphoenolpyruvate and Mg2+ and the concentration of the effector fructose 1,6-bisphosphate. The enzyme was activated by 100 mM-K+ and 32 mM-NH4+ throughout. It was found that an increase in the fructose bisphosphate concentration from 24 microM to 1.2 mM brings about a transition from a sigmoidal to a non-inflected form in the relationships v = f([phosphoenolpyruvate]) and v = f([Mg2+]) together with a large increase in the affinity of these substrates for the enzyme. The binding behaviour of ADP is barely affected by the same change in effector concentration. By contrast, increase in fructose bisphosphate concentration below 24 microM increases the affinity of the enzyme for all its substrates and the sigmoidicity of the corresponding velocity-substrate-concentration relationships. As a result of this change in behaviour it has been found impossible to represent all the data by the exponential model for a regulatory enzyme, and it is suggested (supported by comparisons with previous work) that the failure may reflect a secondary action of the effector upon the enzyme.

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Section: Modelling the Datasupporting

confidence: 88%

Section: Modelling the Datasupporting

confidence: 56%

Section: Modelling the Datamentioning

confidence: 85%

See 1 more Smart Citation

The regulatory properties of yeast pyruvate kinase. Effect of fructose 1,6-bisphosphate

Morris¹,

Ainsworth²,

Kinderlerer³

1986

Biochemical Journal

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“…The fluorescence of tryptophan residues within protein molecules is very sensitive to changes in environment and has been used to monitor subtle conformational changes within proteins (Waldeman et al, 1988). The tryptophanyl fluorescence quench exhibited by yeast pyruvate kinase on addition of Fru(1,6)P2 is profound and has been used successfully to monitor the binding of this allosteric effector to the enzyme (Kuczenski andSuelter, 1970, 1971).…”

Section: Discussionmentioning

confidence: 99%

The cooperative binding of fructose-1,6-bisphosphate to yeast pyruvate kinase.

1992

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The cooperative binding of the allosteric activator fructose‐1,6‐bisphosphate [Fru(1,6)P2] to yeast pyruvate kinase was investigated by equilibrium dialysis and fluorescence quench titration. The results show that yeast pyruvate kinase binds four molecules of Fru(1,6)P2 per tetramer and the observed fluorescence quench follows the binding of the ligand and not the cooperative T to R state transition. Additionally it is shown that the binding of Fru(1,6)P2 to yeast pyruvate kinase is compatible with the model of cooperativity that has been proposed and incorporates an intermediate state, R′, with properties between those of the T and R states.

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“…These included Na+, K+, Tris, and tetramethylammonium (TMA+). The latter two are generally nonfunctional in those enzyme systems that require K+ or Na+ for activity or conformational stability (Evans and Sorger, 1966; Kuczenski and Suelter, 1971), thus allowing for the maintenance of constant ionic strength without interfering with or mimicking specific K+ or Na+ effects.…”

mentioning

confidence: 99%

Monovalent Cations and Striatal Tyrosine Hydroxylase

Kuczenski

1982

Journal of Neurochemistry

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The kinetic properties of soluble tyrosine hydroxylase from rat striatum and the activation of the enzyme by the polyanion heparin were assessed as a function of the monovalent cations K+, Na+, tetramethylammonium (TMA+), and Tris. Substitution of K+ or Na+ for TMA+ or Tris can alter the kinetic properties of tyrosine hydroxylase in the absence of heparin the nature of the interaction of the enzyme with heparin and also the kinetic properties of the heparin-activated enzyme. The data suggest that monovalent cations can support unique conformational states of the enzyme.

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Interactions of fructose 1,6-diphosphate, substrates, and monovalent cations with yeast pyruvate kinase monitored by changes in enzyme fluorescence

Abstract: SUELTERas attempting to distinguish between the substrate and coenzyme sites, and the essential and nonessential tyrosyl residues.

Cited by 25 publications

References 23 publications

The regulatory properties of yeast pyruvate kinase. Effect of fructose 1,6-bisphosphate

The regulatory properties of yeast pyruvate kinase. Effect of fructose 1,6-bisphosphate

The cooperative binding of fructose-1,6-bisphosphate to yeast pyruvate kinase.

Monovalent Cations and Striatal Tyrosine Hydroxylase

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