Information on EC 1.3.1.12 - prephenate dehydrogenase

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The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota

EC NUMBER
COMMENTARY
1.3.1.12
-
RECOMMENDED NAME
GeneOntology No.
prephenate dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
prephenate + NAD+ = 4-hydroxyphenylpyruvate + CO2 + NADH
show the reaction diagram
-
-
-
-
prephenate + NAD+ = 4-hydroxyphenylpyruvate + CO2 + NADH
show the reaction diagram
the T-protein is a bifunctional enzyme showing chorismate mutase, EC 5.4.99.5, and prephenate dehydrogenase activities at two separate active sites
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
decarboxylation
-
-
-
-
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
3-dimethylallyl-4-hydroxybenzoate biosynthesis
-
tyrosine biosynthesis I
-
Phenylalanine, tyrosine and tryptophan biosynthesis
-
Novobiocin biosynthesis
-
Metabolic pathways
-
Biosynthesis of secondary metabolites
-
SYSTEMATIC NAME
IUBMB Comments
prephenate:NAD+ oxidoreductase (decarboxylating)
This enzyme in the enteric bacteria also possesses chorismate mutase activity (EC 5.4.99.5 chorismate mutase) and converts chorismate into prephenate.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
AroQ
O30012
gene name. Trifunctional enzyme EC 5.4.99.5 (chorismate mutase)/EC 4.2.1.51 (prephenate dehydratase)/EC 1.3.1.12 (prephenate dehydrogenase)
bifunctional T-protein
-
-
bifunctional T-protein
Escherichia coli K12
-
-
-
bifunctional T-protein
-
-
chorismate mutase-prephenate dehydratase
-
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme
-
both reactions occur at one active site; complex with EC 5.4.99.5
chorismate mutase-prephenate dehydrogenase bifunctional enzyme
-
complex with EC 5.4.99.5
chorismate mutase-prephenate dehydrogenase bifunctional enzyme
-
complex with EC 5.4.99.5
chorismate mutase-prephenate dehydrogenase bifunctional enzyme
Escherichia coli K12
-
complex with EC 5.4.99.5; complex with EC 5.4.99.5
-
chorismate mutase-T:prephenate dehydrogenase bifunctional enzyme
-
-
chorismate mutase-T:prephenate dehydrogenase bifunctional enzyme
Escherichia coli K12
-
-
-
chorismate mutase-T:prephenate dehydrogenase bifunctional enzyme
-
-
Chorismate mutase/prephenate dehydratase
-
-
Chorismate mutase/prephenate dehydratase
Escherichia coli K12
-
-
-
CM-TyrAp
-
-
CM/PDT/PDHG
O30012
trifunctional enzyme EC 5.4.99.5 (chorismate mutase)/EC 4.2.1.51 (prephenate dehydratase)/EC 1.3.1.12 (prephenate dehydrogenase)
dehydrogenase, prephenate
-
-
-
-
hydroxyphenylpyruvate synthase
-
-
-
-
PDH
-
-
-
-
PDH
Escherichia coli K-12 MG1655
-
-
-
pdhE-1
Soil bacterium
J9XQS6
-
tyrA
Escherichia coli K-12 MG1655, Escherichia coli K12
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9044-92-2
-, prephenate dehydrogenase
94859-19-5
hydroxyphenylpyruvate synthase: chorismate mutase-prephenate dehydrogenase bifunctional enzyme
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Acholeplasma laidlawii JA1
JA1
-
-
Manually annotated by BRENDA team
strain VF5
-
-
Manually annotated by BRENDA team
Archaeoglobus fulgidus DSM 4340
-
SwissProt
Manually annotated by BRENDA team
basonym Alcaligenes eutrophus
-
-
Manually annotated by BRENDA team
K12; strains B and W
-
-
Manually annotated by BRENDA team
L-tyrosine overproducer strain
-
-
Manually annotated by BRENDA team
strain K-12 MG1655
-
-
Manually annotated by BRENDA team
strain K12
-
-
Manually annotated by BRENDA team
Escherichia coli K-12 MG1655
strain K-12 MG1655
-
-
Manually annotated by BRENDA team
Escherichia coli K12
K12
-
-
Manually annotated by BRENDA team
Escherichia coli K12
strain K12
-
-
Manually annotated by BRENDA team
Neurospora sp.
pt MN64
-
-
Manually annotated by BRENDA team
no activity in Actinomycetales
-
-
-
Manually annotated by BRENDA team
no activity in coryneform bacteria
-
-
-
Manually annotated by BRENDA team
Soil bacterium
uncultured alkaline-polluted bacterium
UniProt
Manually annotated by BRENDA team
Streptococcus mutans ATCC 700610
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
metabolism
-, O30012
the enzyme is involved in aromatic amino acid biosynthesis
metabolism
-, P43902
chorismate mutase/prephenate dehydrogenase from Haemophilus influenzae Rd KW20 is a bifunctional enzyme that catalyzes the rearrangement of chorismate to prephenate and the NAD(P)+-dependent oxidative decarboxylation of prephenate to 4-hydroxyphenylpyruvate in tyrosine biosynthesis
metabolism
Archaeoglobus fulgidus DSM 4340
-
the enzyme is involved in aromatic amino acid biosynthesis
-
metabolism
-
chorismate mutase/prephenate dehydrogenase from Haemophilus influenzae Rd KW20 is a bifunctional enzyme that catalyzes the rearrangement of chorismate to prephenate and the NAD(P)+-dependent oxidative decarboxylation of prephenate to 4-hydroxyphenylpyruvate in tyrosine biosynthesis
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
L-arogenate + NAD+
L-tyrosine + NADH + CO2
show the reaction diagram
-
-
-
-
?
L-arogenate + NAD+
L-tyrosine + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
-
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
-
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
Neurospora sp.
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
calorimetric and equilibrium measurements
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
mechanism, kinetic studies
-
-
-
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
mechanism, kinetic studies
-
-
ir
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
mechanism, kinetic studies
-
-
-
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
mechanism, kinetic studies
-
-
ir
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
-
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
ir
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
-
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
ir
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
Neurospora sp.
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
Acholeplasma laidlawii JA1
-
-, biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
Escherichia coli K-12 MG1655
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
Escherichia coli K12
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
Escherichia coli K12
-
-, biosynthesis of L-tyrosine
-
-
-
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
Escherichia coli K12
-
-, biosynthesis of L-tyrosine
-
-
-
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH
show the reaction diagram
-
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH
show the reaction diagram
O67636
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH
show the reaction diagram
-
second step in the biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH
show the reaction diagram
-, O30012
the enzyme is involved in aromatic amino acid biosynthesis
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH
show the reaction diagram
-, O30012
activity with NADP+ as coenzyme is about 10% of that with NAD+, suggesting that NAD+ is likely the preferred and physiological coenzyme
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH + H+
show the reaction diagram
-, P43902
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH + H+
show the reaction diagram
-, P43902
key active-site residues are located at the domain interface, including His200, Arg297 and Ser179, that are involved in catalysis and/or ligand binding and are highly conserved in TyrA proteins
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH + H+
show the reaction diagram
-, P43902
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH + H+
show the reaction diagram
-, P43902
key active-site residues are located at the domain interface, including His200, Arg297 and Ser179, that are involved in catalysis and/or ligand binding and are highly conserved in TyrA proteins
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + H+ + CO2
show the reaction diagram
Soil bacterium
J9XQS6
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + H+ + CO2
show the reaction diagram
Streptococcus mutans, Streptococcus mutans ATCC 700610
-
-
-
-
?
prephenate + NADP+
4-hydroxyphenylpyruvate + NADPH + CO2
show the reaction diagram
-
-
-
-
?
L-arogenate + NAD+
L-tyrosine + NADH + CO2
show the reaction diagram
Escherichia coli K12
-
-
-
-
?
additional information
?
-
-, P43902
a dimeric enzyme, with each monomer consisting of an N-terminal alpha/beta dinucleotide-binding domain and a C-terminal alpha-helical dimerization domain. Absence of an alpha/beta motif in HinfPDH that is present in other TyrA proteins. Residues from this motif are involved in discrimination between NADP+ and NAD+. The loop between beta5 and beta6 in the N-terminal domain is much shorter in HinfPDH and an extra helix is present at the C-terminus. Furthermore, HinfPDH adopts a more closed conformation compared with TyrA proteins that do not have tyrosine bound. This conformational change brings the substrate, cofactor and active-site residues into close proximity for catalysis. An ionic network consisting of Arg297, a key residue for tyrosine binding, a water molecule, Asp206, from the loop between beta5 and beta6, and Arg365', from the additional C-terminal helix of the adjacent monomer, is observed that might be involved in gating the active site. Active site structure, overview
-
-
-
additional information
?
-
-, P43902
a dimeric enzyme, with each monomer consisting of an N-terminal alpha/beta dinucleotide-binding domain and a C-terminal alpha-helical dimerization domain. Absence of an alpha/beta motif in HinfPDH that is present in other TyrA proteins. Residues from this motif are involved in discrimination between NADP+ and NAD+. The loop between beta5 and beta6 in the N-terminal domain is much shorter in HinfPDH and an extra helix is present at the C-terminus. Furthermore, HinfPDH adopts a more closed conformation compared with TyrA proteins that do not have tyrosine bound. This conformational change brings the substrate, cofactor and active-site residues into close proximity for catalysis. An ionic network consisting of Arg297, a key residue for tyrosine binding, a water molecule, Asp206, from the loop between beta5 and beta6, and Arg365', from the additional C-terminal helix of the adjacent monomer, is observed that might be involved in gating the active site. Active site structure, overview
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
-
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
ir
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
-
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
ir
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
Neurospora sp.
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH
show the reaction diagram
-
second step in the biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH
show the reaction diagram
-, O30012
the enzyme is involved in aromatic amino acid biosynthesis
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH + H+
show the reaction diagram
-, P43902
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + H+ + CO2
show the reaction diagram
Soil bacterium
J9XQS6
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
Acholeplasma laidlawii JA1
-
biosynthesis of L-tyrosine
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + CO2 + NADH + H+
show the reaction diagram
-, P43902
-
-
-
?
prephenate + NAD+
4-hydroxyphenylpyruvate + NADH + CO2
show the reaction diagram
Escherichia coli K12
-
biosynthesis of L-tyrosine
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
NAD+
-, O30012
activity with NADP+ as coenzyme is about 10% of that with NAD+, suggesting that NAD+ is likely the preferred and physiological coenzyme
NAD+
-, P43902
binding mode
NADP+
-
slight activity
NADP+
-
NADP+ preferred
additional information
-
NADP+ cannot substitute for NAD+
-
additional information
-
NADP+ cannot substitute for NAD+
-
additional information
-
NADP+ cannot substitute for NAD+
-
additional information
-
NADP+ cannot substitute for NAD+
-
additional information
-
NADP+ cannot substitute for NAD+
-
additional information
-
NADP+ cannot substitute for NAD+
-
additional information
-
NADP+ cannot substitute for NAD+
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Cu2+
Soil bacterium
J9XQS6
0.5 mM, 56.8% residual activity
Fe3+
Soil bacterium
J9XQS6
0.5 mM, 58.7% residual activity
KCl
-
required for optimal activity
Mn2+
Soil bacterium
J9XQS6
0.5 mM, 72% residual activity
SDS
Soil bacterium
J9XQS6
0.10%, 18.3% residual activity
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-hydroxyphenylacetate
-
-
4-hydroxymercuribenzoate
-
-
4-hydroxymercuribenzoate
-
-
4-hydroxyphenylpyruvate
-
-
4-hydroxyphenylpyruvate
-
-
4-hydroxyphenylpyruvate
-
-
4-hydroxyphenylpyruvate
-
-
4-hydroxyphenylpyruvate
-
-
5,5'-dithiobis(2-nitrobenzoate)
-
-
5,6-dihydro-5,6-dihydroxychorismate
-
-
adamantane analogues
-
-
-
adamantane-1,3-diacetate
-
-
adamantane-1-acetate
-
-
adamantane-1-carboxylate
-
-
adamantane-1-phosphonate
-
-
alpha-Methyl-D,L-tyrosine
-
-
chorismate analogues
-
-
-
chorismate-5,6-epoxide
-
-
Co2+
-
weakly inhibits
coumaric acid
-
-
Diethylmalonate
-
-
dihydroxyphenylalanine
-
-
iodoacetamide
-
modification of only one cysteinyl residue results in the inactivation
L-Tyr
-
L-Tyr inhibits TyrAp activity by 90% at concentrations higher than 0.5 mM
L-tyrosine
-
-
L-tyrosine
-
not inhibiting
L-tyrosine
Neurospora sp.
-
-
L-tyrosine
-
feed-back inhibition of both enzyme activities
L-tyrosine
-
Tyr263 or residues 354 to 357 of the enzyme are involved in the feedback inhibition
L-tyrosine
-, O30012
0.5 mM, 50% inhibition
L-tyrosine
-
0.1 mM, less than 20% residual activity
N-benzenesulfonyl-L-phenylalanine
-
-
-
N-Benzyloxycarbonyl-L-phenylalanine
-
-
N-bromosuccinimide
-
-
N-ethylmaleimide
-
-
N-Toluene-p-sulfonyl-L-p-aminophenylalanine
-
-
N-Toluene-p-sulfonyl-L-phenylalanine
-
-
NADH
-
non-competitive inhibition
Tetranitromethane
-
-
trans-2,3-pleiadanedicarboxylic acid
-
i.e. diacid, 50% inhibition at 0.25 mM, inhibits competitively the prephenate dehydrogenase activity, but not the chorismate mutase activity of the enzyme
tyrosine
-
feedback-inhibitor of activity over a wide temperature range, enhances the cooperativity between subunits in the binding of prephenate
tyrosine
-, O67636
binds directly to the active site of the enzyme and not to an allosteric site. Linear competitive inhibition
tyrosine
-, P43902
is bound directly at the catalytic site as a competitive inhibitor
additional information
-
substrate analogues
-
additional information
-
EDTA, MgCl2 and ZnCl2 show no effect on enzyme activity
-
additional information
-, O30012
phenylalanine (up to 5 mM) has no effect. Not inhibited by prephenate or NAD+ at concentrations up to 10 and 2 mM, respectively
-
additional information
Soil bacterium
J9XQS6
not inhibitory: tyrosine
-
additional information
-
not inhibitory: NaCl up to 10 mM, EDTA and dithiothreitol up to 1 mM
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Adamantane 1-acetate
-
-
Bovine serum albumin
-
at low concentrations of prephenate
-
Bovine serum albumin
-
formation of an enzyme-albumin complex with MW of 150000
-
Bovine serum albumin
-
-
-
Guanidine-HCl
-
activates at low concentrations, at higher concentrations concomitant loss of activity with a multi-state pathway of denaturation
L-tryptophan
-
-
L-tryptophan analogues
-
methylated or fluorinated
-
phenylalanine
Neurospora sp.
-
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.017
-
L-arogenate
-
at 30C, wild-type
0.041
-
L-arogenate
-
at 30C, delta19PD
0.097
-
L-arogenate
-
at 55C, wild-type
0.135
-
L-arogenate
-
at 55C, delta19PD
0.136
-
L-arogenate
-
at 80C, wild-type
0.01
-
NAD+
-, O67636
mutant H217A, pH 7.5, 55C
0.012
-
NAD+
-, O67636
mutant H217N, pH 7.5, 55C
0.025
-
NAD+
-
at 30C, wild-type
0.047
-
NAD+
-, O30012
pH 6.5, 87C
0.053
-
NAD+
-
at 30C, delta19PD
0.071
-
NAD+
-
at 55C, delta19PD
0.071
-
NAD+
-, O67636
wild-type, pH 7.5, 55C
0.072
-
NAD+
-
at 55C, wild-type
0.084
-
NAD+
-
at 80C, delta19PD
0.089
-
NAD+
-, O67636
mutant R250Q, pH 7.5, 55C
0.099
-
NAD+
-, O67636
mutant S126A, pH 7.5, 55C
0.103
-
NAD+
-
wild-type enzyme
0.109
-
NAD+
-
at 80C, wild-type
0.128
0.141
NAD+
-
mutant forms H197N and K37Q
0.13
0.15
NAD+
-
-
0.13
0.15
NAD+
-
-
0.13
0.15
NAD+
-
-
0.33
-
NAD+
-
-
0.38
-
NAD+
-
mutant form R294Q
3.7
-
NAD+
-
pH 6.8, 37C
0.017
-
prephenate
-
at 30C, wild-type
0.041
-
prephenate
-
at 30C, delta19PD
0.045
0.09
prephenate
-
-
0.045
0.09
prephenate
-
-
0.045
0.09
prephenate
-
-
0.045
0.09
prephenate
-
-
0.045
0.09
prephenate
-
-
0.045
0.09
prephenate
-
-
0.054
0.055
prephenate
-
mutant forms K37Q and H197N
0.097
-
prephenate
-
at 55C, wild-type
0.104
-
prephenate
-, O67636
mutant H147N, pH 7.5, 55C
0.11
-
prephenate
-
presence of bovine serum albumin
0.135
-
prephenate
-
at 55C, delta19PD
0.135
-
prephenate
-, O67636
wild-type, pH 7.5, 55C
0.136
-
prephenate
-
at 80C, wild-type
0.22
-
prephenate
-
-
0.249
-
prephenate
-
at 80C, delta19PD
0.35
0.37
prephenate
-
-
0.35
0.37
prephenate
-
-
0.45
-
prephenate
-, O30012
pH 6.5, 87C
0.56
-
prephenate
-
cofactor NADP+
0.57
-
prephenate
-
cofactor NAD+
0.75
-
prephenate
-
pH 6.8, 37C
0.83
-
prephenate
-
-
0.87
-
prephenate
Soil bacterium
J9XQS6
pH 8.0, 45C
1.185
-
prephenate
-, O67636
mutant R250Q, pH 7.5, 55C
1.335
-
prephenate
-, O67636
mutant S126A, pH 7.5, 55C
3.213
-
prephenate
-, O67636
mutant H217N, pH 7.5, 55C
4.132
-
prephenate
-, O67636
mutant H217A, pH 7.5, 55C
5
-
prephenate
-
mutant form R294Q
0.249
-
L-arogenate
-
at 80C, delta19PD
additional information
-
additional information
-
Km of enzyme fragments
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.3
-
L-arogenate
-
at 30C, wild-type
2.2
-
L-arogenate
-
at 30C, delta19PD
3
6
L-arogenate
-
at 80C, wild-type
6.8
-
L-arogenate
-
at 55C, wild-type
13
-
L-arogenate
-
at 55C, delta19PD
68
-
L-arogenate
-
at 80C, delta19PD
0.0000583
-
NAD+
-
mutant enzyme H197N
0.3
-
NAD+
-, O67636
mutant H217N, pH 7.5, 55C
0.367
-
NAD+
-
mutant enzyme K37Q
0.417
-
NAD+
-
mutant form R294Q
0.45
-
NAD+
-
wild-type enzyme
0.6
-
NAD+
-, O67636
mutant H217A, pH 7.5, 55C
0.8
-
NAD+
-, O67636
mutant S126A, pH 7.5, 55C
1.1
-
NAD+
-
at 30C, wild-type
1.5
-
NAD+
-
at 30C, delta19PD
6.7
-
NAD+
-
at 55C, wild-type
7.4
-
NAD+
-
pH 6.8, 37C
11.5
-
NAD+
-
at 55C, delta19PD
11.5
-
NAD+
-, O67636
wild-type, pH 7.5, 55C
11.6
-
NAD+
-, O67636
mutant R250Q, pH 7.5, 55C
34
-
NAD+
-
at 80C, wild-type
62
-
NAD+
-
at 80C, delta19PD
0.00005
-
prephenate
-
mutant enzyme H197N
0.0034
-
prephenate
-, O67636
mutant H147N, pH 7.5, 55C
0.383
-
prephenate
-
mutant enzyme K37Q
0.417
-
prephenate
-
mutant form R294Q
0.45
-
prephenate
-
wild-type enzyme
0.5
-
prephenate
-, O67636
mutant H217N, pH 7.5, 55C
0.8
-
prephenate
-, O67636
mutant H217A, pH 7.5, 55C; mutant S126A, pH 7.5, 55C
1.3
-
prephenate
-
at 30C, wild-type
2.2
-
prephenate
-
at 30C, delta19PD
3
6
prephenate
-
at 80C, wild-type
6.8
-
prephenate
-
at 55C, wild-type
9.9
-
prephenate
-, O67636
mutant R250Q, pH 7.5, 55C
10.08
-
prephenate
Soil bacterium
J9XQS6
pH 8.0, 45C
10.5
-
prephenate
-
pH 6.8, 37C
13
-
prephenate
-
at 55C, delta19PD
13
-
prephenate
-, O67636
wild-type, pH 7.5, 55C
68
-
prephenate
-
at 80C, delta19PD
135
-
prephenate
-
-
170.4
-
prephenate
-
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2
-
NAD+
-
pH 6.8, 37C
7
11.6
-
prephenate
Soil bacterium
J9XQS6
pH 8.0, 45C
353
13.9
-
prephenate
-
pH 6.8, 37C
353
880
-
prephenate
-, O30012
pH 6,5, 87C
353
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.17
-
2-hydroxyphenylacetate
-
pH 7.8, 37C
0.13
-
4-hydroxyphenylpyruvate
-
-
0.4
-
4-hydroxyphenylpyruvate
-
-
0.11
-
5,6-dihydro-5,6-dihydroxychorismate
-
pH 7.8, 37C
0.15
-
adamantane-1,3-diacetate
-
pH 7.8, 37C
0.13
-
adamantane-1-acetate
-
pH 7.8, 37C
0.31
-
adamantane-1-carboxylate
-
pH 7.8, 37C
0.14
-
adamantane-1-phosphonate
-
pH 7.8, 37C
0.13
-
chorismate-5,6-epoxide
-
pH 7.8, 37C
0.2
-
coumaric acid
-
-
1.3
-
D-tyrosine
-
-
0.47
-
dihydroxyphenylalanine
-
-
0.3
-
L-phenylalanine
-
-
0.7
-
L-tryptophan
-
-
0.003
-
L-tyrosine
-
-
0.017
-
L-tyrosine
-
with prephenate as variable substrate
0.02
-
L-tyrosine
-
with NAD+ as variable substrate
0.035
-
L-tyrosine
-
-
0.06
-
L-tyrosine
-
-
5.5
-
L-tyrosine
-
-
0.21
-
trans-2,3-pleiadanedicarboxylic acid
-
pH 7.8, 37C
0.0159
-
tyrosine
-, O67636
wild-type, pH 7.5, 55C
0.037
-
tyrosine
-, O67636
mutant S126A, pH 7.5, 55C
0.37
-
tyrosine
-, O67636
mutant R250Q, pH 7.5, 55C
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.01
-
NaCl
-
IC50: 0.01 mM
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
12.6
-
-
pH 6.8, 37C
25.2
-
-
purified recombinant enzyme fragment comprising amino acid residues 93-373
33
-
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme
55
-
-
purified recombinant enzyme fragment comprising amino acid residues 96-373
98
-
-
purified recombinant wild-type enzyme
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
assay method
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
activity of enzyme fragments
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
-
-
about, assay at
7.5
-
-
at 55C
7.8
-
-
assay at
8
-
Soil bacterium
J9XQS6
-
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
8.5
Soil bacterium
J9XQS6
more than 75% of maximum activity within
8
-
-
50% of maximum activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
40
-
assay at 37C
37
-
-
assay at
45
-
Soil bacterium
J9XQS6
-
87
-
-, O30012
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
55
-
not active at 65C
35
50
Soil bacterium
J9XQS6
more than 80% of maximum activity within
37
-
-
decrease of activity above
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
-
Soil bacterium
J9XQS6
-
PDB
SCOP
CATH
ORGANISM
Aquifex aeolicus (strain VF5)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Streptococcus mutans serotype c (strain ATCC 700610 / UA159)
Streptococcus thermophilus (strain ATCC BAA-250 / LMG 18311)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
31000
-
-
sequence analysis
36420
-
-
MALDI-TOF studies of the recombinant His-rMtPDH
42040
-
-
calculated from nucleotide sequence
52500
-
-
calculated from nucleotide sequence
55000
65000
-
gel filtration, gel electrophoresis, sucrose density gradient centrifugation
66000
-
-
gel filtration
68000
-
-
gel filtration
75000
-
-
gel filtration
76000
81000
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme, gel filtration, gel electrophoresis
78000
100000
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme, sedimentation equilibrium centrifugation
78000
100000
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme, gel electrophoresis
78000
100000
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme, gel electrophoresis; chorismate mutase-prephenate dehydrogenase bifunctional enzyme, sedimentation equilibrium centrifugation
78000
100000
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme, gel electrophoresis; chorismate mutase-prephenate dehydrogenase bifunctional enzyme, gel filtration; chorismate mutase-prephenate dehydrogenase bifunctional enzyme, sedimentation equilibrium centrifugation
78000
100000
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme, gel electrophoresis; chorismate mutase-prephenate dehydrogenase bifunctional enzyme, gel filtration
140000
-
-
gel filtration
150000
-
-
gel filtation
420000
-
-, O30012
gel filtration
additional information
-
-
formation of higher MW aggregates in presence of tyrosine and NAD+
additional information
-
-
-
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
Soil bacterium
J9XQS6
x * 31400, calculated
dimer
-
2 * 39000-42000, SDS-PAGE
dimer
-
2 * 39000-42000, SDS-PAGE
dimer
-
wild-type full length enzyme and some enzyme fragments form homodimers
dimer
-
gel filtration
dimer
-, P43902
SDS-PAGE and gel filtration, the dimeric enzyme, with each monomer consisting of an N-terminal alpha/beta dinucleotide-binding domain and a C-terminal alpha-helical dimerization domain, structure comparisons, overview
dimer
-
2 * 41800, calculated, 2 * 41000, SDS-PAGE
dimer
-
SDS-PAGE and gel filtration, the dimeric enzyme, with each monomer consisting of an N-terminal alpha/beta dinucleotide-binding domain and a C-terminal alpha-helical dimerization domain, structure comparisons, overview
-
dimer
Streptococcus mutans ATCC 700610
-
2 * 41800, calculated, 2 * 41000, SDS-PAGE
-
heterodimer
-
1 * 37000 + 1 * 34000, SDS-PAGE
homohexamer
-, O30012
6 * 66000, SDS-PAGE; 6 * 70946, calculated from sequence
additional information
-
MW of recombinant enzyme fragments, overview, structural localization of the different enzyme activities on the enzyme molecule
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
proteolytic digestion of recombinant wild-type enzyme by papain, pH 6.8, 37C
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
in complex with NAD+, by hanging drop vapor diffusion technique at room temperature, one modified nucleotide-binding domain and a novel helical prephenate binding domain, active site formed at the domain interface and shared between the subunits of the dimer, access to active site may be regulated via a gated mechanism, modulated by an ionic network involving a conserved arginine, active site residues include Ser126 and Lys246 and the catalytic His147, two functional domains of chorismate mutase-prephenate dehydrogenase are interdependent
-
prephenate dehydrogenase bound with NAD+ plus either 4-hydroxyphenylpyuvate, 4-hydroxyphenylpropionate, or L-tyrosine. Resiudes His147 and Arg250 are key catalytic and binding groups, respectively, and Ser126 participates in both catalysis and substrate binding through the ligand 4-hydroxyl group. Inhibitor tyrosine binds directly to the active site of the enzyme and not to an allosteric site
-, O67636
prephenate dehydrogenase component of the TyrA protein from strain Rd KW20 in complex with inhibitor tyrosine and cofactor NAD+, sitting drop vapour diffusion method, 200 nl of 19.6 mg/ml protein in 20 mM HEPES pH 8.0, 200 mM NaCl, 40 mM imidazole, 1 mM TCEP are mixed with 200 nl reservoir solution containing 0.04 M potassium dihydrogen phosphate, 20.0% v/v glycerol and 16.0% w/v PEG 8000, X-ray diffrraction structure determination and analysis at 2.0 A resolution
-, P43902
to 2.1 A resolution. The N-terminal alpha/beta domain has a Rossman fold for binding a NAD+ molecule. The C-terminal domain adopts a helical architecture and is involved in homo-dimerization. NAD+ binding stabilizes the active site and facilitates the substrate, prephenate, binding
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
dithiothreitol, protection against thermal inactivation
additional information
-
-
L-tyrosine, protection against thermal inactivation
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
dialysis inactivates
-
glycerol stabilizes
-
glycerol stabilizes
-
stability in Tris-HCl buffer higher than in phosphate buffer
-
tyrosine stabilizes
-
bovine serum albumin, bovine casein or prephenate stabilizes
-
citrate stabilizes
-
dialysis inactivates
-
dithiothreitol stabilizes
-
EDTA stabilizes
-
glycerol stabilizes
-
NAD+ plus tyrosine stabilizes
-
purified enzyme and diluted solution is unstable
-
stability in Tris-HCl buffer higher than in phosphate buffer
-
thiols stabilize
-
tyrosine stabilizes
-
unstable in buffers of low ionic strength lacking glycerol
-
unstable in crude extracts
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, 50 mM phosphate buffer, pH 7.4, at least 0.7 mg/ml protein, 10 months
-
-15C, 0.02% sodium azide, N2-atmosphere, more than 0.1 mg/ml protein
-
0-4C, 0.1 M Tris-HCl buffer, pH 7.5, 1 mM EDTA, 1 mM DTT, 1 mM tyrosine, 70% w/v ammonium sulfate, for at least 10 days
-
-20C, 0.1 M N-ethylmorpholine, pH 7.0, 21 mM citrate, 10% v/v glycerol, 1 mM EDTA, 1 mM DTT
-
-20C, 0.1 M Tris-HCl buffer, pH 7.5, 1 mM EDTA, 1 mM DTT, 1 mM tyrosine, 70% w/v ammonium sulfate, several weeks
-
-20C, 50 mM Cl/N-ethylmorpholine pH 7.2, 1 mM EDTA, 1 mM sodium citrate, 0.02% w/v sodium azide, 50% v/v glycerol, 10 mM DTT, N2-atmosphere, at least 3 months
-
-80C, purified recombinant His-tagged wild-type enzyme, complete loss of activity when stored in 0.1 M sodium citrate, pH 7.5, 10% glycerol, 1 mM DTT, or moderate loss in activity when stored in 0.1 M MES, pH 7.5, 0.051 M N-ethylmorpholine, 0.01 M diethanolamine, 1 mM EDTA, 1 mM DTT, 10% glycerol,
-
4C, 0.1 M N-ethylmorpholine, pH 7.0, 21 mM citrate, 10% v/v glycerol, 1 mM EDTA, 1 mM DTT, up to 4 weeks
-
4C, 0.1 M phosphate buffer pH 7.6, over-night, complete loss of activity
-
4C, ammonium sulfate precipitate
-
4C, -20C, or -80C, 50 mM Tris, 25% glycerol, no significant differences in activity, even after one month of storage
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
by Ni-NTA column
-
by nickel-affinity chromatography
-
copurification with arogenate dehydrogenase; partial
-
not associated with other enzymatic activities
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme; one-step procedure by affinity chromatography
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme; partial
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme; wild-type and mutant enzyme
-
His6-tagged proteins, to homogeneity by gel filtration
-
recombinant His-tagged wild-type enzyme and enzyme fragments
-
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography, cleavage of the tag by TEV protease
-, P43902
chorismate mutase-prephenate dehydrogenase bifunctional enzyme; partial
-
together with arogenate dehydrogenase
-
His-rMtPDH, to near homogeneity by Ni-NTA affinity chromatography, ca. 94% pure
-
copurification with pretyrosine dehydrogenase
-
copurification with arogenate dehydrogenase
-
recombinant enzyme
Soil bacterium
J9XQS6
recombinant protein
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
into vector pET-15b, expression as His-tagged protein in Escherichia coli
-
into vector pET15b and expressed in Escherichia coli BL21-Gold (DE3)
-
overexpressed in Escherichia coli
-, O30012
gene tyrA, genetic mapping of chorismate mutase and prephenate dehydrogenase activity, expression of the wild-type enzyme, the 2 activity domains, and gene fragments exhibiting either chorismate mutase or prephenate dehydrogenase activity as His-tagged proteins and peptides in strain BL21(DE3), respectively
-
genetic separability of chorismate mutase and prephenate dehydrogenase in Escherichia coli
-
mutants cloned into pZE21-MCS1 and transformed to Escherichia coli DH5alpha, wild-type and mutants subcloned into vector pET-30XA/LIC and expressed in Escherichia coli BL21(DE3)
-
overexpression
-
T7 expression system
-
expression of His-tagged enzyme in Escherichia coli
-, P43902
into tyrA-pET30a and expressed in Escherichia coli BL21(DE3) as His-rMtPDH
-
monofunctional prephenate dehydrogenase, expressed in Escherichia coli
-
expression in Escherichia coli
Soil bacterium
J9XQS6
expression in Escherichia coli
Q8DUW0
expressed in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
DELTA1-19
-
delta19PD, more sensitive to temperature effects yielding a half-life of 55 min at 95C versus 2 h for PD
H147N
-, O67636
inactive, binds prephenate with apparent affinity similar to wild-type
H217A
-, O67636
40fold increase in Km for prephenate, no inhibition by tyrosine
R250Q
-, O67636
10fold increase in the Km for prephenate and a 20fold increase in Ki for tyrosine
S126A
-, O67636
15fold reduction in kcat, a 10fold increase in the Km value for prephenate, and a 2-fold increase in Ki for tyrosine
H131A
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme, mutant enzyme retains 10% of dehydrogenase activity and 30% mutase activity compared to the wild-type enzyme, His131 is not an essential residue whose protonation state is critical for catalysis or substrate binding
H189N
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme, mutant enzyme shows less than 0.01% the activity of wild-type mutase and dehydrogenase
H197N
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme, mutant enzyme exhibits no significant dehydrogenase activity, retains nearly wild-type mutase activity and unaltered Michaelis constants for chorismate, prephenate and NAD+
K37A
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme, mutant protein poorly expressed, no detectable mutase or dehydrogenase activity, structural changes may result in its inactivity and instability
H217N
-, O67636
30fold increase in Km for prephenate, no inhibition by tyrosine
additional information
-
delta19-PDH, delta25-PDH, and delta28-PDH, which produce stable and soluble proteins, delta36-PDH, delta52-PDH, and delta55-PDH are unstable
K37Q
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme, mutant enzyme exhibits no mutase activity while retaining wild-type dehydrogenase activity
additional information
-
feedback-inhibition-resistant mutants toward L-tyrosine, chorismate mutase activities similar to those of the wild-type, amino acid substitutions either at Tyr263 or at residues 354 to 357
R294Q
-
chorismate mutase-prephenate dehydrogenase bifunctional enzyme, R294Q substitution reduces the affinity of the enzyme for prephenate, Arg294 interacts electrostatically with the ring carboxylate at C1 of prephenate
additional information
Escherichia coli K-12 MG1655
-
feedback-inhibition-resistant mutants toward L-tyrosine, chorismate mutase activities similar to those of the wild-type, amino acid substitutions either at Tyr263 or at residues 354 to 357
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
Mycobacterium tuberculosis PDH is a monofunctional protein, does not possess any chorismate mutase activity unlike many other enteric bacteria