Information on EC 1.3.1.12 - prephenate dehydrogenase

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

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
-
Biosynthesis of secondary metabolites
-
Metabolic pathways
-
Novobiocin biosynthesis
-
Phenylalanine, tyrosine and tryptophan biosynthesis
-
tyrosine biosynthesis I
-
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
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
-
-
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
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
-, 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
-
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 + 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 + 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+
-, 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
-
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
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
0.053
-
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
-
-
0.56
-
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
14330
11.6
-
prephenate
Soil bacterium
J9XQS6
pH 8.0, 45C
15738
13.9
-
prephenate
-
pH 6.8, 37C
15738
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
-
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
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
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)
-
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
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genetic separability of chorismate mutase and prephenate dehydrogenase in Escherichia coli
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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)
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overexpression
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T7 expression system
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expression of His-tagged enzyme in Escherichia coli
-, P43902
into tyrA-pET30a and expressed in Escherichia coli BL21(DE3) as His-rMtPDH
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monofunctional prephenate dehydrogenase, expressed in Escherichia coli
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expression in Escherichia coli
Soil bacterium
J9XQS6
expression in Escherichia coli
Q8DUW0
expressed in Escherichia coli
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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
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chorismate mutase-prephenate dehydrogenase bifunctional enzyme, mutant enzyme shows less than 0.01% the activity of wild-type mutase and dehydrogenase
H197N
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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
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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
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delta19-PDH, delta25-PDH, and delta28-PDH, which produce stable and soluble proteins, delta36-PDH, delta52-PDH, and delta55-PDH are unstable
K37Q
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chorismate mutase-prephenate dehydrogenase bifunctional enzyme, mutant enzyme exhibits no mutase activity while retaining wild-type dehydrogenase activity
additional information
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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
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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
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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
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APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
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Mycobacterium tuberculosis PDH is a monofunctional protein, does not possess any chorismate mutase activity unlike many other enteric bacteria