|
Plant Physiol, October 1999, Vol. 121, pp. 589-598
Plant Succinic Semialdehyde Dehydrogenase. Cloning, Purification,
Localization in Mitochondria, and Regulation by Adenine
Nucleotides1
Karin B.
Busch and
Hillel
Fromm2*
Department of Plant Sciences, The Weizmann Institute of Science,
Rehovot 76100, Israel
Succinic
semialdehyde dehydrogenase (SSADH) is one of three enzymes constituting
the  -aminobutyric acid shunt. We have cloned the cDNA for SSADH from
Arabidopsis, which we designated SSADH1. SSADH1 cDNA encodes a protein
of 528 amino acids (56 kD) with high similarity to SSADH from
Escherichia coli and human (>59% identity). A sequence
similar to a mitochondrial protease cleavage site is present 33 amino
acids from the N terminus, indicating that the mature mitochondrial
protein may contain 495 amino acids (53 kD). The native recombinant
enzyme and the plant mitochondrial protein have a tetrameric molecular
mass of 197 kD. Fractionation of plant mitochondria revealed its
localization in the matrix. The purified recombinant enzyme showed
maximal activity at pH 9.0 to 9.5, was specific for succinic
semialdehyde (K0.5 = 15 µM), and exclusively used NAD+ as a cofactor
(Km = 130 ± 77 µM).
NADH was a competitive inhibitor with respect to NAD+
(Ki = 122 ± 86 µM).
AMP, ADP, and ATP inhibited the activity of SSADH
(Ki = 2.5-8 mM). The
mechanism of inhibition was competitive for AMP, noncompetitive for
ATP, and mixed competitive for ADP with respect to NAD+.
Plant SSADH may be responsive to mitochondrial energy charge and
reducing potential in controlling metabolism of -aminobutyric acid.
1
This work was supported by a grant (to H.F.)
from the Leo and Julia Forchheimer Center for Molecular Genetics,
Weizmann Institute of Science. K.B. was the recipient of a MINERVA
postdoctoral fellowship.
2
Present address: Centre for Plant Sciences,
Leeds Institute for Plant Biotechnology and Agriculture, The University
of Leeds, Leeds LS2 9JT, UK.
*
Corresponding author; e-mail bgyhf{at}leeds.ac.uk; fax
44-113-233-3144.
© 1999 American Society of Plant Physiologists
This article has been cited by other articles:
|
|
|
|
 
C. Jaeger, A. Gessler, S. Biller, H. Rennenberg, and J. Kreuzwieser
Differences in C metabolism of ash species and provenances as a consequence of root oxygen deprivation by waterlogging
J. Exp. Bot.,
November 1, 2009;
60(15):
4335 - 4345.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. M. Clark, R. Di Leo, O. R. Van Cauwenberghe, R. T. Mullen, and B. J. Shelp
Subcellular localization and expression of multiple tomato {gamma}-aminobutyrate transaminases that utilize both pyruvate and glyoxylate
J. Exp. Bot.,
July 1, 2009;
60(11):
3255 - 3267.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. M. Clark, R. Di Leo, P. K. Dhanoa, O. R. Van Cauwenberghe, R. T. Mullen, and B. J. Shelp
Biochemical characterization, mitochondrial localization, expression, and potential functions for an Arabidopsis {gamma}-aminobutyrate transaminase that utilizes both pyruvate and glyoxylate
J. Exp. Bot.,
April 1, 2009;
60(6):
1743 - 1757.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Gao, W. Xiong, X.-b. Li, C.-F. Gao, Y.-l. Zhang, H. Li, and Q.-y. Wu
Identification of the proteomic changes in Synechocystis sp. PCC 6803 following prolonged UV-B irradiation
J. Exp. Bot.,
March 4, 2009;
(2009)
ern356v1.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. L. Allan, J. P. Simpson, S. M. Clark, and B. J. Shelp
{gamma}-Hydroxybutyrate accumulation in Arabidopsis and tobacco plants is a general response to abiotic stress: putative regulation by redox balance and glyoxylate reductase isoforms
J. Exp. Bot.,
June 1, 2008;
59(9):
2555 - 2564.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Studart-Guimaraes, A. Fait, A. Nunes-Nesi, F. Carrari, B. Usadel, and A. R. Fernie
Reduced Expression of Succinyl-Coenzyme A Ligase Can Be Compensated for by Up-Regulation of the {gamma}-Aminobutyrate Shunt in Illuminated Tomato Leaves
Plant Physiology,
November 1, 2007;
145(3):
626 - 639.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Mazzucotelli, A. Tartari, L. Cattivelli, and G. Forlani
Metabolism of {gamma}-aminobutyric acid during cold acclimation and freezing and its relationship to frost tolerance in barley and wheat
J. Exp. Bot.,
November 1, 2006;
57(14):
3755 - 3766.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Watanabe, T. Kodaki, and K. Makino
A Novel {alpha}-Ketoglutaric Semialdehyde Dehydrogenase: EVOLUTIONARY INSIGHT INTO AN ALTERNATIVE PATHWAY OF BACTERIAL L-ARABINOSE METABOLISM
J. Biol. Chem.,
September 29, 2006;
281(39):
28876 - 28888.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. E. Breitkreuz, W. L. Allan, O. R. Van Cauwenberghe, C. Jakobs, D. Talibi, B. Andre, and B. J. Shelp
A Novel {gamma}-Hydroxybutyrate Dehydrogenase: IDENTIFICATION AND EXPRESSION OF AN ARABIDOPSIS cDNA AND POTENTIAL ROLE UNDER OXYGEN DEFICIENCY
J. Biol. Chem.,
October 17, 2003;
278(42):
41552 - 41556.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Bouche, A. Fait, D. Bouchez, S. G. Moller, and H. Fromm
Mitochondrial succinic-semialdehyde dehydrogenase of the {gamma}-aminobutyrate shunt is required to restrict levels of reactive oxygen intermediates in plants
PNAS,
May 27, 2003;
100(11):
6843 - 6848.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. H. Millar, L. J. Sweetlove, P. Giege, and C. J. Leaver
Analysis of the Arabidopsis Mitochondrial Proteome
Plant Physiology,
December 1, 2001;
127(4):
1711 - 1727.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. T. Coleman, T. K. Fang, S. A. Rovinsky, F. J. Turano, and W. S. Moye-Rowley
Expression of a Glutamate Decarboxylase Homologue Is Required for Normal Oxidative Stress Tolerance in Saccharomyces cerevisiae
J. Biol. Chem.,
January 5, 2001;
276(1):
244 - 250.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
