The Stereochemistry of the Reaction Catalyzed by D-Glyceric 3-Dehydrogenase<sup>*</sup>

Krakow, Gladys; Udaka, Shigezo; Vennesland, Birgit

doi:10.1021/bi00908a010

Cited by 20 publications

(8 citation statements)

References 18 publications

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“…Tartre activity increased during incubation in propionatetartronic ser mineral salts media. This increase in enzyme activ- (96,107).m ity occurred before the initiation of exponential grown ongl growth and probably represents enzyme derepresreferred to sion due to protein turnover, for synthesis These data agree with the concept that E-26V on this subsi possesses a selective advantage during adaptation The propc to propionate owing to its capacity to form a late carbolij higher activity of a-hydroxyglutarate synthase. It is suggests It is proposed that the a-hydroxyglutarate pathform hydrox way is physiologically significant in E. coli E-26V a second mc as a mechanism for the formation Of C4 acids semnialdehydi directly from propionate.…”

Section: (214)supporting

confidence: 82%

Alternate pathways of metabolism of short-chain fatty acids.

Wegener¹,

Reeves²,

Rabin³

et al. 1968

Bacteriological Reviews

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Section: (214)supporting

confidence: 82%

Alternate pathways of metabolism of short-chain fatty acids.

Wegener¹,

Reeves²,

Rabin³

et al. 1968

Bacteriological Reviews

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“…The retention of tritium is generally larger than similar retention of deuterium in previous experiments. This probably reflects the larger discrimination effects expected with the heavier isotope, as previously explained (Krakow et al, 1962).…”

Section: Table I Stereospecificitysupporting

confidence: 63%

“…For these measurements, about 1.5 mg of the pyridine nucleotides were employed, and sufficient unlabeled carrier nicotinamide was added to dilute the labeled nicotinamide about 1700-fold. The counting procedure was the same as previously described (Krakow et al, 1962). The quenching effect of IV'-methylnicotinamide iodide on the scintillation count was high and no effort was made to obtain accurate specific activity measurements for this derivative, but the other compounds could be counted in the range of 1-5 mg per sample with only a small correction for quenching by the 2-pyridone.…”

Section: Methodsmentioning

confidence: 99%

“…The ether was removed by evaporation, and the nicotinamide was recrystallized twice from benzene, mp 130-131°. Two samples of nicotinamide (about 2 and 4 mg) were weighed out and counted in a scintillation counter as previously described (Krakow et al 1962). For comparison, the tritium content of the DPN (T) + prior to reduction and reoxidation was determined similarly after dilution with unlabeled nicotinamide.…”

Section: Methodsmentioning

confidence: 99%

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Some Stereospecificity Studies with Tritiated Pyridine Nucleotides^*

Krakow¹,

Ludowieg²,

Mather³

et al. 1963

Biochemistry

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The stereospecificity of hydrogen transfer for the pyridine nucleotide has been ascertained for a number of hitherto uninvestigated dehydrogenase reactions. These determinations have been made with the help of appropriate reference enzymes, and of DPN and TPN labeled with tritium at the 4 position of the nicotinamide ring. Evidence is presented that the following enzymes use the same position (i.e., A or a) as yeast alcohol dehydrogenase: DPN-linked formic dehydrogenase from pea seeds, DPN-linked glyoxylic reductase from spinach leaves, TPN-linked glyoxylic reductase from pea seeds, TPN-linked dihydroorotic dehydrogenase from an aerobic bacterium, and TPN-linked malic enzyme from liver. The opposite stereospecificity (B or ß) is demonstrated for the DPN-linked liver dehydrogenase which oxidizes uridine diphosphoglucose to uridine diphosphoglucuronic acid.

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“…The midpoint potential of the orotate/DHO couple is -258 mV (25). We determined a value for the two electron potential of the ligand-free enzyme of -310 mV.…”

Section: Discussionmentioning

confidence: 99%

Insight into the Chemistry of Flavin Reduction and Oxidation inEscherichia coliDihydroorotate Dehydrogenase Obtained by Rapid Reaction Studies

2001

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Dihydroorotate dehydrogenase (DHOD) oxidizes dihydroorotate (DHO) to orotate in the only redox reaction of pyrimidine biosynthesis. The enzyme from Escherichia coli is a membrane-bound FMN-containing enzyme that is thought to use ubiquinone as the oxidizing substrate. The chemistry of the reduction of the flavin in DHOD from E. coli by the substrate dihydroorotate (DHO) was studied at 4 degrees C in anaerobic stopped-flow experiments conducted over a broad range of pH values. A Michaelis complex that was characterized by a approximately 20 nm red-shift of the oxidized flavin absorbance formed within the dead-time of the stopped-flow instrument ( approximately 1 ms) upon mixing with DHO. The flavin of the intermediate was reduced by DHO, forming a reduced flavin-orotate charge-transfer complex. The rate constant for the flavin reduction reaction increased with pH, from a value of 1 s(-1) at pH 6.5 to approximately 360 s(-1) at pH values greater than an observed pK(a) of 9.5 which was ascribed to Ser175, the active-site base. At all pH values, the reduced flavin-orotate charge-transfer complex dissociated too slowly to be catalytically relevant. Therefore, the oxidizing quinone substrate must bind to the reduced enzyme-orotate complex at a site distinct from the substrate binding site, in agreement with steady-state kinetic studies [Björnberg, O., Grüner, A.-C., Roepstorff, P., and Jensen, K. F. (1999) Biochemistry 38, 2899-2908]. Menadione was used as a model quinone substrate to oxidize dithionite-reduced DHOD. The reduced enzyme-orotate complex reacted rapidly with menadione (180 s(-1)), demonstrating that the reduced enzyme-orotate complex is a catalytically competent intermediate.

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The Stereochemistry of the Reaction Catalyzed by D-Glyceric 3-Dehydrogenase^*

Cited by 20 publications

References 18 publications

Alternate pathways of metabolism of short-chain fatty acids.

Alternate pathways of metabolism of short-chain fatty acids.

Some Stereospecificity Studies with Tritiated Pyridine Nucleotides^*

Insight into the Chemistry of Flavin Reduction and Oxidation inEscherichia coliDihydroorotate Dehydrogenase Obtained by Rapid Reaction Studies

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The Stereochemistry of the Reaction Catalyzed by D-Glyceric 3-Dehydrogenase*

Cited by 20 publications

References 18 publications

Alternate pathways of metabolism of short-chain fatty acids.

Alternate pathways of metabolism of short-chain fatty acids.

Some Stereospecificity Studies with Tritiated Pyridine Nucleotides*

Insight into the Chemistry of Flavin Reduction and Oxidation inEscherichia coliDihydroorotate Dehydrogenase Obtained by Rapid Reaction Studies

Contact Info

Product

Resources

About

The Stereochemistry of the Reaction Catalyzed by D-Glyceric 3-Dehydrogenase^*

Some Stereospecificity Studies with Tritiated Pyridine Nucleotides^*