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Originally published In Press as doi:10.1074/jbc.M205466200 on October 21, 2002
J. Biol. Chem., Vol. 277, Issue 51, 49655-49661, December 20, 2002
The Activity of the Arabidopsis Bifunctional
Lysine-ketoglutarate Reductase/Saccharopine Dehydrogenase Enzyme of
Lysine Catabolism Is Regulated by Functional Interaction between
Its Two Enzyme Domains*
Xiaohong
Zhu,
Guiliang
Tang, and
Gad
Galili
From the Department of Plant Sciences, The Weizmann Institute of
Science, Rehovot 76100, Israel
Lysine-ketoglutarate
reductase/saccharopine dehydrogenase (LKR/SDH) is a bifunctional enzyme
catalyzing the first two steps of lysine catabolism in animals and
plants. To elucidate the biochemical signification of the linkage
between the two enzymes of LKR/SDH, namely lysine ketoglutarate and
saccharopine dehydrogenase, we employed various truncated and mutated
Arabidopsis LKR/SDH polypeptides expressed in yeast.
Activity analyses of the different recombinant polypeptides under
conditions of varying NaCl levels implied that LKR, but not SDH
activity, is regulated by functional interaction between the LKR and
SDH domains, which is mediated by the structural conformation of the
linker region connecting them. Because LKR activity of plant LKR/SDH
enzymes is also regulated by casein kinase 2 phosphorylation, we
searched for such potential regulatory phosphorylation sites using
matrix-assisted laser desorption/ionization time-of-flight mass
spectrometry and site-directed mutagenesis. This analysis identified
Ser-458 as a candidate for this function. We also tested a hypothesis
suggesting that an EF-hand-like sequence at the C-terminal part of the
LKR domain functions in a calcium-dependent assembly of
LKR/SDH into a homodimer. We found that this region is essential for
LKR activity but that it does not control a
calcium-dependent assembly of LKR/SDH. The relevance of our
results to the in vivo function of LKR/SDH in lysine
catabolism in plants is discussed. In addition, because the linker
region between LKR and SDH exists only in plants but not in animal
LKR/SDH enzymes, our results suggest that the regulatory properties of
LKR/SDH and, hence, the regulation of lysine catabolism are different
between plants and animals.
*
This work was supported by grants from the Israel
Academy of Sciences and Humanities, National Council for Research and
Development, Israel and by the MINERVA Foundation, Germany.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
An incumbent of the Bronfman Chair of Plant Sciences. To
whom correspondence should be addressed. Tel.: 972-8-9343511; Fax: 972-8-9344181; E-mail: gad.galili@weizmann.ac.il.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.

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Copyright © 2002 by the American Society for Biochemistry and Molecular Biology.
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