Catalytically active filaments – pyruvate decarboxylase from Neurospora crassa. pH-controlled oligomer structure and catalytic function

Authors

  • Stefanie Hüttl,

    1.  Division of Enzymology, Institute of Biochemistry & Biotechnology, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany
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    • Present addresses
      Division of Molecular Plant Physiology and Biophysics, Botany I, Julius-von-Sachs-Institute of Biological Sciences, Julius-Maximilian-University Würzburg, Würzburg, Germany

  • Juliane Fiebig,

    1.  Division of Enzymology, Institute of Biochemistry & Biotechnology, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany
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    • Present addresses
      Division of Molecular Plant Physiology and Biophysics, Botany I, Julius-von-Sachs-Institute of Biological Sciences, Julius-Maximilian-University Würzburg, Würzburg, Germany

  • Steffen Kutter,

    1.  Division of Enzymology, Institute of Biochemistry & Biotechnology, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany
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    • Kern Laboratory, Howard Hughes Medical Institute and Department of Biochemistry, Brandeis University, Waltham, MA, USA

  • Gerd Hause,

    1.  Microscopy Unit, Biocenter of the Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany
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  • Hauke Lilie,

    1.  Division of Technical Biochemistry, Institute of Biochemistry & Biotechnology, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany
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  • Michael Spinka,

    1.  Division of Enzymology, Institute of Biochemistry & Biotechnology, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany
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  • Stephan König

    1.  Division of Enzymology, Institute of Biochemistry & Biotechnology, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany
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  • Note
    Stefanie Hüttl and Juliane Fiebig contributed equally to this work

S. König, Abteilung Enzymologie, Institut für Biochemie/Biotechnologie, Martin-Luther-Universität Halle-Wittenberg, Kurt-Mothes-Str. 3, 06120 Halle (Saale), Germany
Fax: +49 345 55 24829
Tel: +49 345 55 27014
E-mail: stephan.koenig@biochemtech.uni-halle.de

Abstract

Pyruvate decarboxylase is a key enzyme in organisms whose energy metabolism is based on alcoholic fermentation. The enzyme catalyses the nonoxidative decarboxylation of 2-oxo acids in the presence of the cofactors thiamine diphosphate and magnesium ions. Pyruvate decarboxylase species from yeasts and plant seeds studied to date are allosterically activated by their substrate pyruvate. However, detailed kinetic studies on the enzyme from Neurospora crassa demonstrate for the first time the lack of substrate activation for a yeast pyruvate decarboxylase species. The quaternary structure of this enzyme species is also peculiar because it forms filamentous structures. The complex enzyme structure was analysed using a number of methods, including small-angle X-ray solution scattering, transmission electron microscopy, analytical ultracentrifugation and size-exclusion chromatography. These measurements were complemented by detailed kinetic studies in dependence on the pH.

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