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Information on EC 2.7.2.4 - aspartate kinase and Organism(s) Saccharomyces cerevisiae and UniProt Accession P10869
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2.7.2.4 aspartate kinaseIUBMB Comments
The enzyme from Escherichia coli is a multifunctional protein, which also catalyses the reaction of EC 1.1.1.3 homoserine dehydrogenase. This is also the case for two of the four isoenzymes in Arabidopsis thaliana. The equilibrium constant strongly favours the reaction from right to left, i.e. the non-physiological direction of reaction.
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Word Map
- 2.7.2.4
- corynebacterium
- glutamicum
-
threonine-sensitive
- l-lysine
- l-threonine
- dihydrodipicolinate
-
i-homoserine
- diaminopimelate
- semialdehyde
-
feedback-resistant
-
aspartate-derived
-
feedback-insensitive
- aec
- l-isoleucine
- s-2-aminoethyl-l-cysteine
-
lysine-producing
- flavum
- thialysine
- meso-diaminopimelate
- ectoine
- synthesis
- agriculture
- medicine
- nutrition
- food industry
The taxonomic range for the selected organisms is: Saccharomyces cerevisiae
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
aspartokinase, aspartate kinase, thra1, aspartokinase ii, aspartokinase iii, aspartokinase i, akiii, thra2, ak iii, ak ii, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
AK
aspartate kinase (phosphorylating)
-
-
-
-
aspartic kinase
-
-
-
-
aspartokinase
beta-aspartokinase
-
-
-
-
AK
-
-
-
-
aspartokinase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
phospho group transfer
-
-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
KEGG
-
-, -, -, -, -, -, -, -, -, -, -, -, -, -
SYSTEMATIC NAME
IUBMB Comments
ATP:L-aspartate 4-phosphotransferase
The enzyme from Escherichia coli is a multifunctional protein, which also catalyses the reaction of EC 1.1.1.3 homoserine dehydrogenase. This is also the case for two of the four isoenzymes in Arabidopsis thaliana. The equilibrium constant strongly favours the reaction from right to left, i.e. the non-physiological direction of reaction.
CAS REGISTRY NUMBER
COMMENTARY
9012-50-4
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + L-aspartate
ADP + 4-phospho-L-aspartate
-
first step in the branched pathway leading to synthesis of threonine and methionine from aspartate
-
r
ATP + L-aspartate
ADP + phospho-L-aspartate
-
-
-
-
?
additional information
?
-
-
D-aspartate, L-glutamate and beta-alanine are inactive as substitutes for L-aspartate in the forward reaction, in the reverse reaction ADP cannot be replaced by AMP, UDP, GDP or IDP
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + L-aspartate
ADP + 4-phospho-L-aspartate
-
first step in the branched pathway leading to synthesis of threonine and methionine from aspartate
-
r
ATP + L-aspartate
ADP + phospho-L-aspartate
-
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Fe2+
Mg2+
Mn2+
Fe2+
-
can partially replace Mg2+, but to a smaller extent than Mn2+
Mg2+
-
essential for enzyme activity, can be partially replaced by Mn2+ and to a smaller extent by Fe2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
L-threonine
75% inhibition at 10 mM, 22°C culture temperature, 22°C assay temperature, 71% inhibition at 10 mM, 37°C culture temperature, 22°C assay temperature, 87% inhibition at 10 mM, 22°C culture temperature, 37°C assay temperature, 86% inhibition at 10 mM, 37°C culture temperature, 37°C assay temperature, 93% inhibition of glutathione-S-transferase fusion protein at 10 mM, 22°C assay temperature, 73% inhibition of glutathione-S-transferase fusion protein at 10 mM, 37°C assay temperature
L-threonine
-
allosteric inhibition, causes a decrease in the apparent molecular mass of the enzyme, no inhibition is detected in the threonine insensitive mutant up to 50 mM threonine
L-threonine
reduced sensitivity to threonine inhibition, 0% inhibition at 10 mM, 22°C culture temperature, 22°C assay temperature, 15% inhibition at 10 mM, 37°C culture temperature, 22°C assay temperature, 4% inhibition at 10 mM, 22°C culture temperature, 37°C assay temperature, 14% inhibition at 10 mM, 37°C culture temperature, 37°C assay temperature
L-threonine
resistance to feedback inhibition, 16% inhibition at 10 mM, 22°C culture temperature, 22°C assay temperature, 15% inhibition at 10 mM, 37°C culture temperature, 22°C assay temperature, 9% inhibition at 10 mM, 22°C culture temperature, 37°C assay temperature, 6% inhibition at 10 mM, 37°C culture temperature, 37°C assay temperature, 31% inhibition of GST-fusion protein at 10 mM, 22°C assay temperature, 16% inhibition of glutathione-S-transferase fusion protein at 10 mM, 37°C assay temperature
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.2
ATP
-
glutathione-S-transferase fusion protein, in the presence of 10 mM aspartate
1.2
ATP
-
threonine insensitive mutant, in the presence of 10 mM aspartate
1.1
L-aspartate
-
threonine insensitive mutant, in the presence of 10 mM ATP
1.2
L-aspartate
-
glutathione-S-transferase fusion protein, in the presence of 10 mM ATP
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.4
L-threonine
-
glutathione-S-transferase fusion protein, in the presence of 10 mM aspartate and 10 mM ATP
1.4
L-threonine
-
wildtype, in the presence of 10 mM aspartate and 10 mM ATP, binds threonine in a hyperbolic manner
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
10.6
glutathione-S-transferase fusion protein, 37°C assay temperature
5.6
glutathione-S-transferase fusion protein, 22°C assay temperature
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
280000
-
glutathione-S-transferase fusion protein, in the presence of threonine, gel filtration
297000
-
glutathione-S-transferase fusion protein, in the presence of threonine, blue native gel electrophoresis
344000
345000
346000
344000
-
threonine insensitive mutant, with and without threonine in the buffer, blue native gel electrophoresis
345000
-
threonine insensitive mutant, with and without threonine in the buffer, gel filtration
346000
-
glutathione-S-transferase fusion protein, blue native gel electrophoresis
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pentamer
-
glutathione-S-transferase fusion protein, in the presence of threonine, gel filtration
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A406T
-
site-directed mutagenesis, 30fold more strongly inhibited by threonine
E254A
-
9.1fold decrease in kcat for aspartate and 11fold decrease in kcat for ATP
E279A
-
kcat is decreased 47 times for aspartate and 44 times for ATP
G25D
-
site-directed mutagenesis, reduced affinity for its substrates aspartate and ATP
H292A
-
4.5 times increase in Km for ATP and 120 times decrease in kcat for ATP
K18R
-
Km values for aspartate similar to wildtype, Km value for ATP 2.9fold decreased
K26I
-
site-directed mutagenesis, reduced affinity for its substrates aspartate and ATP
R419A
-
10fold decrease in kcat for aspartate and 8.9fold decrease in kcat for ATP
S23A
-
differs significantly only in the kcat/Km ratio, which is decreased 4fold for aspartate and 3.7fold for ATP
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
partial
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli and as fusion protein with GST (glutathione-S-transferase) in Saccharomyces cerevisiae
-
single aspartate kinase encoded by the HOM3 gene, Escherichia coli DH5alpha as host
-
the gene is cloned into the NheI and BamHI restriction enzyme sites of the expression vector pET28 to create pET28+AKSc, expression in Escherichia coli
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Black, S.
Conversion of aspartic acid to homoserine
Methods Enzymol.
5
820-827
1962
Saccharomyces cerevisiae
-
Truffa-Bachi, P.
Microbial aspartokinases
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
8
509-553
1973
Azotobacter sp., Bacillus cereus, Bacillus licheniformis, Bacillus subtilis, Cereibacter sphaeroides, Corynebacterium glutamicum, Escherichia coli, Geobacillus stearothermophilus, Neurospora crassa, no activity in Edwardsiella sp., no activity in Providencia sp., Paenibacillus polymyxa, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas putida, Rhodobacter capsulatus, Rhodocyclus tenuis, Rhodospirillum rubrum, Saccharomyces cerevisiae, Salmonella enterica subsp. enterica serovar Typhimurium, [Brevibacterium] flavum
-
Arevalo-Rodriguez, M.; Calderon, I.L.; Holmberg, S.
Mutations that cause threonine sensitivity identify catalytic and regulatory regions of the aspartate kinase of Saccharomyces cerevisiae
Yeast
15
1331-1345
1999
Saccharomyces cerevisiae
Bareich, D.C.; Wright, G.D.
Functionally important amino acids in Saccharomyces cerevisiae aspartate kinase
Biochem. Biophys. Res. Commun.
311
597-603
2003
Saccharomyces cerevisiae
Marina, P.; Martinez-Costa, O.H.; Calderon, I.L.; Aragon, J.J.
Characterization of the aspartate kinase from Saccharomyces cerevisiae and of its interaction with threonine
Biochem. Biophys. Res. Commun.
321
584-591
2004
Saccharomyces cerevisiae
Velasco, I.; Arevalo-Rodriguez, M.; Marina, P.; Calderon, I.L.
A new mutation in the yeast aspartate kinase induces threonine accumulation in a temperature-regulated way
Yeast
22
99-110
2005
Saccharomyces cerevisiae, Saccharomyces cerevisiae (P10869), Saccharomyces cerevisiae HT1(pIVUts31d), Saccharomyces cerevisiae SG211
Lo, C.C.; Bonner, C.A.; Xie, G.; DSouza, M.; Jensen, R.A.
Cohesion group approach for evolutionary analysis of aspartokinase, an enzyme that feeds a branched network of many biochemical pathways
Microbiol. Mol. Biol. Rev.
73
594-651
2009
Aquifex aeolicus, Bdellovibrio bacteriovorus, Bdellovibrio bacteriovorus HD100, Chlamydia sp., Corynebacterium glutamicum (P26512), Escherichia coli, Francisella tularensis subsp. novicida, Francisella tularensis subsp. novicida U112, Leptospira borgpetersenii, Leptospira interrogans, Maricaulis maris, Maricaulis maris MCS10, Methanopyrus kandleri, Myxococcus xanthus, Pyrococcus furiosus, Saccharomyces cerevisiae, Thermotoga maritima, Thermotoga petrophila
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