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Activation of thiamin diphosphate in enzymes

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1998

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Hübner, Gerhard
Tittmann, Kai
Killenberg-Jabs, Margrit
Schäffner, Jörg
Spinka, Michael
Neef, Holger
Kern, Dorothee
Kern, Gunther
Schneider, Gunter
Wikner, Christer

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Biochimica et Biophysica Acta / Protein Structure and Molecular Enzymology. 1998, 1385(2), pp. 221-228. ISSN 0167-4838. Available under: doi: 10.1016/S0167-4838(98)00070-3

Zusammenfassung

Activation of the coenzyme ThDP was studied by measuring the kinetics of deprotonation at the C2 carbon of thiamin diphosphate in the enzymes pyruvate decarboxylase, transketolase, pyruvate dehydrogenase complex, pyruvate oxidase, in site-specific mutant enzymes and in enzyme complexes containing coenzyme analogues by proton/deuterium exchange detected by 1H-NMR spectroscopy. The respective deprotonation rate constant is above the catalytic constant in all enzymes investigated. The fast deprotonation requires the presence of an activator in pyruvate decarboxylase from yeast, showing the allosteric regulation of this enzyme to be accomplished by an increase in the C2-H dissociation rate of the enzyme-bound thiamin diphosphate. The data of the thiamin diphosphate analogues and of the mutant enzymes show the N1′ atom and the 4′-NH2 group to be essential for the activation of the coenzyme and a conserved glutamate involved in the proton abstraction mechanism of the enzyme-bound thiamin diphosphate.

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570 Biowissenschaften, Biologie

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Pyruvate decarboxylase, Pyruvate dehydrogenase multienzyme complex, Pyruvate oxidase, Regulation, Thiamin diphosphate, Transketolase

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ISO 690HÜBNER, Gerhard, Kai TITTMANN, Margrit KILLENBERG-JABS, Jörg SCHÄFFNER, Michael SPINKA, Holger NEEF, Dorothee KERN, Gunther KERN, Gunter SCHNEIDER, Christer WIKNER, Sandro GHISLA, 1998. Activation of thiamin diphosphate in enzymes. In: Biochimica et Biophysica Acta / Protein Structure and Molecular Enzymology. 1998, 1385(2), pp. 221-228. ISSN 0167-4838. Available under: doi: 10.1016/S0167-4838(98)00070-3
BibTex
@article{Hubner1998Activ-6571,
  year={1998},
  doi={10.1016/S0167-4838(98)00070-3},
  title={Activation of thiamin diphosphate in enzymes},
  number={2},
  volume={1385},
  issn={0167-4838},
  journal={Biochimica et Biophysica Acta / Protein Structure and Molecular Enzymology},
  pages={221--228},
  author={Hübner, Gerhard and Tittmann, Kai and Killenberg-Jabs, Margrit and Schäffner, Jörg and Spinka, Michael and Neef, Holger and Kern, Dorothee and Kern, Gunther and Schneider, Gunter and Wikner, Christer and Ghisla, Sandro}
}
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