Article (Scientific journals)
Saccharomyces cerevisiae Forms D-2-Hydroxyglutarate and Couples its Degradation to D-Lactate Formation via a Cytosolic Transhydrogenase.
Becker-Kettern, Julia; Paczia, Nicole; Conrotte, Jean-Francois et al.
2016In The Journal of Biological Chemistry
Peer reviewed
 

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Keywords :
2-hydroxyglutarate; dehydrogenase; enzyme kinetics; flavoprotein; inborn error of metabolism; transhydrogenase; yeast metabolism
Abstract :
[en] The D or L form of 2-hydroxyglutarate (2HG) accumulates in certain rare neurometabolic disorders and high D-2HG levels are also found in several types of cancer. Although 2HG has been detected in Saccharomyces cerevisiae, its metabolism in yeast has remained largely unexplored. Here we show that S. cerevisiae actively forms the D enantiomer of 2HG. Accordingly, the S. cerevisiae genome encodes two homologs of the human D-2HG dehydrogenase: Dld2, which, as its human homolog, is a mitochondrial protein, and the cytosolic protein Dld3. Intriguingly, we found that a dld3Delta knockout strain accumulates millimolar levels of D-2HG, while a dld2Delta knockout strain displayed only very moderate increases in D-2HG. Recombinant Dld2 and Dld3, both currently annotated as D-lactate dehydrogenases, efficiently oxidized D-2HG to alpha-ketoglutarate. Depletion of D-lactate levels in the dld3Delta, but not in the dld2Delta mutant, led to the discovery of a new type of enzymatic activity, carried by Dld3, to convert D-2HG to alpha-ketoglutarate, namely an FAD-dependent transhydrogenase activity using pyruvate as a hydrogen acceptor. We also provide evidence that Ser3 and Ser33, which are primarily known for oxidizing 3-phosphoglycerate in the main serine biosynthesis pathway, in addition reduce alpha-ketoglutarate to D-2HG using NADH and represent major intracellular sources of D-2HG in yeast. Based on our observations, we propose that D-2HG is mainly formed and degraded in the cytosol of S. cerevisiae cells in a process that couples D-2HG metabolism to the shuttling of reducing equivalents from cytosolic NADH to the mitochondrial respiratory chain via the D-lactate dehydrogenase Dld1.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Becker-Kettern, Julia ;  University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB)
Paczia, Nicole ;  University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB)
Conrotte, Jean-Francois
Kay, Daniel ;  University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB)
Guignard, Cedric
Jung, Paul ;  University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB)
Linster, Carole  ;  University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB)
External co-authors :
no
Language :
English
Title :
Saccharomyces cerevisiae Forms D-2-Hydroxyglutarate and Couples its Degradation to D-Lactate Formation via a Cytosolic Transhydrogenase.
Publication date :
2016
Journal title :
The Journal of Biological Chemistry
Peer reviewed :
Peer reviewed
Commentary :
Copyright (c) 2016, The American Society for Biochemistry and Molecular Biology.
Available on ORBilu :
since 04 February 2016

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