Inhibition of lactate dehydrogenase by high concentrations of pyruvate: The nature and removal of the inhibitor

Coulson, Christopher J.; Rabin, B. R.

doi:10.1016/0014-5793(69)80171-7

Cited by 44 publications

(14 citation statements)

References 21 publications

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“…\ J Fig.2B shows that the data are compatible with Eqn (6). From the intercept and the slope determined from Fig.…”

Section: Nadh As Variable Substratesupporting

confidence: 61%

“…One of these is based on the formation of an abortive ternary complex involving enzyme, NAD + and pyruvate [l -51. The other has been attributed to the presence in the pyruvate preparations of a contaminating inhibitor or tautomeric isomer end-pyruvate [6,7]. I have observed in the studies of the human erythrocyte glycolysis that the inhibitory effect of pyruvate on lactate dehydrogenase depends on the concentration of NADH cofactor.…”

mentioning

confidence: 99%

“…At low concentration of substrate A, the linear portion of the Lineweaver-Burk plot is, in some cases, difficult to obtain and the measurement of A becomes difficult. Therefore, a choice of optimal concentrations of A and B is necessary for proper measurement of the parameter A .The derivation of the kinetic constants K, and Klb was based on the special case where K, = Ki,; in this case the apparent Michaelis-Menten constant K,' is a linear function of [B] as shown in Eqn(6).…”

mentioning

confidence: 99%

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Inhibition of Human Erythrocyte Lactate Dehydrogenase by High Concentrations of Pyruvate. Evidence for the Competitive Substrate Inhibition

Wang

1977

Eur J Biochem

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The mechanism of the inhibitory effect of high concentrations of pyruvate on human erythrocyte lactate dehydrogenase has been studied by the use of a new parameter, A , defined as the difference between the reciprocals of initial reaction rates obtained from experimental measurements and hypothetical linear Lineweaver-Burk plots. This parameter served as a method for differentiating between the competitive and uncompetitive substrate inhibition. Results of this study indicate that pyruvate is a competitive substrate inhibitor. It is suggested that the inhibitory effect of pyruvate is due to its competition with NADH for binding to the free enzyme and formation of an inactive enzyme . pyruvate binary complex. The competitive nature of pyruvate inhibition is further supported by the results of a kinetic study with NADH as the variable substrate.The dissociation constant of the inactive enzyme . pyruvate binary complex was determined to be 101 pM. The physiological significance of the inhibitory effect could be to preserve a level of NADH concentration necessary for other vital enzymic reactions of living cells despite the presence of a high concentration of pyruvate.Two mechanisms have been proposed to explain the inhibitory effect exhibited by high concentrations of pyruvate on lactate dehydrogenase activity. One of these is based on the formation of an abortive ternary complex involving enzyme, NAD + and pyruvate [l -51. The other has been attributed to the presence in the pyruvate preparations of a contaminating inhibitor or tautomeric isomer end-pyruvate [6,7]. I have observed in the studies of the human erythrocyte glycolysis that the inhibitory effect of pyruvate on lactate dehydrogenase depends on the concentration of NADH cofactor. The inhibitory effect of pyruvate is compatible with the competitive substrate inhibition mechanism as theoretically predicted by Dalziel [S]. Such an inhibitory effect is a specific phenomenon for the ordered multiple-substrate system when a substrate, other than the first substrate, binds to the free enzyme and forms an inactive binary complex. Therefore, in contrast to the proposal that an abortive complex formation or a contaminating inhibitor of the substrate preparation is responsible for inhibition, results of this kinetic

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“…\ J Fig.2B shows that the data are compatible with Eqn (6). From the intercept and the slope determined from Fig.…”

Section: Nadh As Variable Substratesupporting

confidence: 61%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Inhibition of Human Erythrocyte Lactate Dehydrogenase by High Concentrations of Pyruvate. Evidence for the Competitive Substrate Inhibition

Wang

1977

Eur J Biochem

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“…Coulsen and Kabin reported that the reduced pyruvate adduct of ( AcPy)DPN is a far better inhibitor of LDH than the corresponding DPN compound (88). The adducts are competitive with DPNH and noncompetitive with pyruvate, as might be expected from their structural properties.…”

Section: Johannes Evehse and Nathan 0 Kaplanmentioning

confidence: 99%

Lactate Dehydrogenases: Structure and Function

Everse¹,

Kaplan²

1973

Advances in Enzymology - And Related Areas of Molecular Biology

192

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“…This product is an inhibitor of lactate dehydrogenase but because it cyclizes ( Fig. 1) its potency is diminished and it is no more effective at binding to the enzyme than is NAD+ itself (Coulson and Rabin, 1969). Several analogues of NAD' in which -C(0)NH2 is replaced by other groups, including -C(0)CH3 and -C(S)NH2 (Anderson and Kaplan, 1959) and -C(O)H (Kaplan et al, 1956), are substrates.…”

Section: I926mentioning

confidence: 99%

Inhibition of Lactate Production in Rat Brain Extracts and Synaptosomes by 3‐[4‐(Reduced 3‐Pyridine Aldehyde‐Adenine Dinucleotide)]‐Pyruvate

Cooper

Lai

Coleman

et al. 1987

Journal of Neurochemistry

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In basic solutions, pyruvate enolizes and reacts (through its 3-carbon) with the 4-carbon of the nicotinamide ring of NAD+, yielding an NAD-pyruvate adduct in which the nicotinamide ring is in the reduced form. This adduct is a strong inhibitor of lactate dehydrogenase, presumably because it binds simultaneously to the NADH and pyruvate sites. The potency of the inhibition, however, is muted by the adduct's tendency to cyclize to a lactam. We prepared solutions of the pyruvate adduct of NAD+ and of NAD+ analogues in which the -C(O)NH2 of NAD+ was replaced with -C(S)NH2, -C(O)CH3, and -C(O)H. Of the four, only the last analogue, 3-[4-(reduced 3-pyridine aldehyde-adenine dinucleotide)]-pyruvate (RAP) cannot cyclize and it was found to be the most potent inhibitor of beef heart and rat brain lactate dehydrogenases. The inhibitor binds very tightly to the NADH site (Ki approximately 1 nM for the A form). Even at high concentrations (20 microM), RAP had little or no effect on rat brain glyceraldehyde-3-phosphate, pyruvate, alpha-ketoglutarate, isocitrate, soluble and mitochondrial malate, and glutamate dehydrogenases. The glycolytic enzymes, hexokinase and phosphofructokinase, were similarly unaffected. RAP strongly inhibited lactate production from glucose in rat brain extracts but was less effective in inhibiting lactate production from glucose in synaptosomes.

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Inhibition of lactate dehydrogenase by high concentrations of pyruvate: The nature and removal of the inhibitor

Cited by 44 publications

References 21 publications

Inhibition of Human Erythrocyte Lactate Dehydrogenase by High Concentrations of Pyruvate. Evidence for the Competitive Substrate Inhibition

Inhibition of Human Erythrocyte Lactate Dehydrogenase by High Concentrations of Pyruvate. Evidence for the Competitive Substrate Inhibition

Lactate Dehydrogenases: Structure and Function

Inhibition of Lactate Production in Rat Brain Extracts and Synaptosomes by 3‐[4‐(Reduced 3‐Pyridine Aldehyde‐Adenine Dinucleotide)]‐Pyruvate

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