Information on EC 1.14.16.1 - phenylalanine 4-monooxygenase and Organism(s) Homo sapiens

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EC NUMBER
COMMENTARY hide
1.14.16.1
-
RECOMMENDED NAME
GeneOntology No.
phenylalanine 4-monooxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-phenylalanine + tetrahydrobiopterin + O2 = L-tyrosine + 4a-hydroxytetrahydrobiopterin
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
oxygenation
-
S-oxygenation
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
L-phenylalanine degradation I (aerobic)
-
-
L-phenylalanine degradation V
-
-
L-tyrosine biosynthesis IV
-
-
phenylalanine metabolism
-
-
Phenylalanine metabolism
-
-
Phenylalanine, tyrosine and tryptophan biosynthesis
-
-
Folate biosynthesis
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
L-phenylalanine,tetrahydrobiopterin:oxygen oxidoreductase (4-hydroxylating)
The active centre contains mononuclear iron(II). The reaction involves an arene oxide that rearranges to give the phenolic hydroxy group. This results in the hydrogen at C-4 migrating to C-3 and in part being retained. This process is known as the NIH-shift. The 4a-hydroxytetrahydrobiopterin formed can dehydrate to 6,7-dihydrobiopterin, both spontaneously and by the action of EC 4.2.1.96, 4a-hydroxytetrahydrobiopterin dehydratase. The 6,7-dihydrobiopterin can be enzymically reduced back to tetrahydrobiopterin, by EC 1.5.1.34, 6,7-dihydropteridine reductase, or slowly rearranges into the more stable compound 7,8-dihydrobiopterin.
CAS REGISTRY NUMBER
COMMENTARY hide
9029-73-6
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
physiological function
-
key enzyme in the sulfoxidation of S-carboxymethyl-L-cysteine S-oxide and its thioester metabolites S-methyl-L-cysteine, N-acetyl-S-carboxymethyl-L-cysteine, and N-acetyl-S-methyl-L-cysteine
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(S)-butyl-L-cysteine + 5,6,7,8-tetrahydrobiopterin + O2
?
show the reaction diagram
-
low activity
-
-
?
(S)-carboxymethyl-L-cysteine + 5,6,7,8-tetrahydrobiopterin + O2
(S)-carboxymethyl-L-cysteine S-oxide + dihydrobiopterin + H2O
show the reaction diagram
-
low activity
-
-
?
(S)-ethyl-L-cysteine + 5,6,7,8-tetrahydrobiopterin + O2
(S)-ethyl-L-cysteine S-oxide + dihydrobiopterin + H2O
show the reaction diagram
-
-
-
-
?
(S)-ethyl-L-cysteine + 5,6,7,8-tetrahydrobiopterin + O2
?
show the reaction diagram
-
low activity
-
-
?
(S)-methyl-ergothionine + 5,6,7,8-tetrahydrobiopterin + O2
?
show the reaction diagram
-
low activity
-
-
?
(S)-methyl-L-cysteine + 5,6,7,8-tetrahydrobiopterin + O2
(S)-methyl-L-cysteine S-oxide + dihydrobiopterin + H2O
show the reaction diagram
-
low activity
-
-
?
(S)-propyl-L-cysteine + 5,6,7,8-tetrahydrobiopterin + O2
?
show the reaction diagram
-
low activity
-
-
?
3-(2-thienyl)-L-alanine + 6-methyltetrahydropterin + O2
? + 6-methyldihydropterin + H2O
show the reaction diagram
-
-
-
-
?
3-(2-thienyl)-L-alanine + tetrahydrobiopterin + O2
? + dihydrobiopterin + H2O
show the reaction diagram
-
-
-
-
?
4-fluorophenylalanine + tetrahydrobiopterin + O2
?
show the reaction diagram
-
-
-
-
-
L-methionine + 5,6,7,8-tetrahydrobiopterin + O2
?
show the reaction diagram
L-norleucine + tetrahydrobiopterin + O2
? + dihydrobiopterin + H2O
show the reaction diagram
-
5% of the activity with 3-(2-thienyl)-L-alanine
-
-
?
L-Phe + tetrahydrobiopterin + O2
L-tyrosine + dihydrobiopterin + H2O
show the reaction diagram
-
-
-
-
?
L-phenylalanine + (6R)-L-erythro-5,6,7,8-tetrahydrobiopterin + O2
L-tyrosine + 4a-hydroxy-(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin
show the reaction diagram
L-phenylalanine + (6R)-L-erythro-5,6,7,8-tetrahydrobiopterin + O2
L-tyrosine + 4a-hydroxytetrahydrobiopterin
show the reaction diagram
-
-
-
-
?
L-phenylalanine + 5,6,7,8-tetrahydro-L-biopterin + O2
L-tyrosine + 4a-hydroxytetrahydrobiopterin
show the reaction diagram
-
-
-
-
?
L-phenylalanine + 5,6,7,8-tetrahydrobiopterin + O2
L-tyrosine + 4a-hydroxy-tetrahydrobiopterin
show the reaction diagram
-
substrate binding by His285, Trp326, Arg270, Ser349, and Trp278
-
-
?
L-phenylalanine + 5,6,7,8-tetrahydrobiopterin + O2
L-tyrosine + 4a-hydroxytetrahydrobiopterin
show the reaction diagram
L-phenylalanine + 6-methyl-tetrahydrobiopterin + O2
L-tyrosine + 6-methyl-4a-hydroxy-tetrahydrobiopterin
show the reaction diagram
-
-
-
-
?
L-phenylalanine + 6-methyltetrahydropterin + O2
L-tyrosine + 6-methyldihydropterin + H2O
show the reaction diagram
-
-
-
-
?
L-phenylalanine + tetrahydrobiopterin + O2
L-tyrosine + 4a-hydroxy-tetrahydrobiopterin
show the reaction diagram
L-phenylalanine + tetrahydrobiopterin + O2
L-tyrosine + 4a-hydroxytetrahydrobiopterin
show the reaction diagram
L-phenylalanine + tetrahydrobiopterin + O2
L-tyrosine + dihydrobiopterin + H2O
show the reaction diagram
L-thienylalanine + tetrahydrobiopterin + O2
?
show the reaction diagram
-
-
-
-
?
L-tryptophan + tetrahydrobiopterin + O2
?
show the reaction diagram
-
truncated enzyme containing C-terminal 334 amino acids
-
-
?
N-acetyl-(S)-carboxymethyl-L-cysteine + 5,6,7,8-tetrahydro-L-biopterin + O2
N-acetyl-(S)-carboxymethyl-L-cysteine S-oxide + ?
show the reaction diagram
-
-
-
-
?
N-acetyl-(S)-carboxymethyl-L-cysteine + 5,6,7,8-tetrahydrobiopterin + O2
?
show the reaction diagram
-
-
-
-
?
N-acetyl-(S)-methyl-L-cysteine + 5,6,7,8-tetrahydro-L-biopterin + O2
N-acetyl-(S)-methyl-L-cysteine S-oxide + ?
show the reaction diagram
-
-
-
-
?
N-acetyl-(S)-methyl-L-cysteine + 5,6,7,8-tetrahydrobiopterin + O2
?
show the reaction diagram
-
-
-
-
?
N-acetyl-S-carboxymethyl-L-cysteine + O2
?
show the reaction diagram
-
-
-
-
?
N-acetyl-S-methyl-L-cysteine + O2
?
show the reaction diagram
-
-
-
-
?
phenylalanine + tetrahydrobiopterin + O2
tyrosine + 4a-hydroxytetrahydrobiopterin
show the reaction diagram
-
PAH is a key enzyme in the metabolic pathway of phenylalanine. Deficiency in PAH leads to high and persistent levels of this amino acid in theplasma of phenylketonuria patients, causing permanent neurological damage
-
-
ir
S-carboxy-methyl-L-cysteine + 5,6,7,8-tetrahydrobiopterin + O2
S-carboxymethyl-L-cysteine S-oxide + dihydrobiopterin + H2O
show the reaction diagram
-
poor substrate
-
-
?
S-carboxymethyl-L-cysteine + 5,6,7,8-tetrahydro-L-biopterin + O2
S-carboxymethyl-L-cysteine S-oxide + dihydrobiopterin + H2O
show the reaction diagram
-
-
-
-
?
S-carboxymethyl-L-cysteine + 5,6,7,8-tetrahydrobiopterin + O2
S-carboxymethyl-L-cysteine S-oxide + dihydrobiopterin + H2O
show the reaction diagram
-
-
-
-
?
S-carboxymethyl-L-cysteine + tetrahydrobiopterin + O2
?
show the reaction diagram
-
conversion to the (S)-sulfoxide
-
-
?
S-carboxymethyl-L-cysteine + tetrahydrobiopterin + O2
S-carboxymethyl-L-cysteine S-oxide + dihydrobiopterin + H2O
show the reaction diagram
-
-
-
-
?
S-methyl-ergothionine + 5,6,7,8-tetrahydrobiopterin + O2
?
show the reaction diagram
-
-
-
-
?
S-methyl-L-cysteine + 5,6,7,8-tetrahydrobiopterin + O2
S-methyl-L-cysteine S-oxide + dihydrobiopterin + H2O
show the reaction diagram
-
poor substrate
-
-
?
S-methyl-L-cysteine + O2
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
L-phenylalanine + (6R)-L-erythro-5,6,7,8-tetrahydrobiopterin + O2
L-tyrosine + 4a-hydroxy-(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin
show the reaction diagram
-
-
-
-
?
L-phenylalanine + 5,6,7,8-tetrahydrobiopterin + O2
L-tyrosine + 4a-hydroxytetrahydrobiopterin
show the reaction diagram
-
-
-
-
?
L-phenylalanine + tetrahydrobiopterin + O2
L-tyrosine + 4a-hydroxy-tetrahydrobiopterin
show the reaction diagram
L-phenylalanine + tetrahydrobiopterin + O2
L-tyrosine + 4a-hydroxytetrahydrobiopterin
show the reaction diagram
phenylalanine + tetrahydrobiopterin + O2
tyrosine + 4a-hydroxytetrahydrobiopterin
show the reaction diagram
-
PAH is a key enzyme in the metabolic pathway of phenylalanine. Deficiency in PAH leads to high and persistent levels of this amino acid in theplasma of phenylketonuria patients, causing permanent neurological damage
-
-
ir
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin
(7R)-tetrahydrobiopterin
-
8fold lower affinity and activity compared with 6(R)BH4
(7S)-tetrahydrobiopterin
-
8fold lower affinity and activity compared with 6(R)BH4
6-methyl-tetrahydrobiopterin
-
-
tetrahydrobiopterin
additional information
-
cofactor binding analysis, overview
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cl-
-
bound by residue S391
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(7R)-5,6,7,8-tetrahydrobiopterin
-
-
(7R)-5,6,7,8-tetrahydropterin
-
0.001 mM, 50% inhibition at 0.5 mM phenylalanine, 0.004 mM, 50% inhibition at 0.1 mM phenylalanine, recombinant enzyme
-
(7R)-tetrahydrobiopterin
-
slight inhibition, synthetic pathway, overview, conformational structure by NMR
(7S)-tetrahydrobiopterin
-
strong, competitive inhibition, synthetic pathway, overview, conformational structure by NMR
2,2'-dipyridyl
-
99.0% inhibition at 1.0 mM using L-phenylalanine as substrate, 99.0% inhibition at 1.0 mM using S-carboxymethyl-L-cysteine as substrate
3-iodotyrosine
-
3.0% inhibition at 1.0 mM using L-phenylalanine as substrate, 5.2% inhibition at 1.0 mM using S-carboxymethyl-L-cysteine as substrate
4-chloromercuribenzoate
-
1 mM, complete inhibition after 10 min
4-Chlorophenylalanine
4-Fluorophenylalanine
-
above 1 mM
6-Fluorotryptophan
-
2.5% inhibition at 1.0 mM using L-phenylalanine as substrate, 4.5% inhibition at 1.0 mM using S-carboxymethyl-L-cysteine as substrate
7(S)-tetrahydrobiopterin
-
-
Copper-chelating agents
-
-
-
diethyldithiocarbamate
-
1 mM, 58% inhibition
DL-alpha-tocopherol
-
strong inhibition
Iron-chelating agents
-
-
-
L-methionine
-
competitive inhibitor of the C-oxidation of L-phenylalanine
L-phenylalanine
-
competitive inhibition of the S-oxidation of S-carboxymethyl-L-cysteine, 92.8% inhibition at 1.0 mM using S-carboxymethyl-L-cysteine as substrate
S-carboxy-methyl-L-cysteine
-
competitive inhibitor of the C-oxidation of L-phenylalanine
S-carboxymethyl-L-cysteine
-
competitive inhibitor of the aromatic C-oxidation of L-phenylalanine, 25.2% inhibition at 5.0 mM using L-phenylalanine as substrate
S-methyl-ergothionine
-
competitive inhibitor of the C-oxidation of L-phenylalanine
S-methyl-L-cysteine
-
competitive inhibitor of the C-oxidation of L-phenylalanine
tetrahydrobiopterin
-
excessive dosages of BH4 inhibit PAH under normal phenylalanine conditions in vivo and activate PAH under conditions of high phenylalanine, overview
Thiol-binding reagents
-
-
-
tryptophan
-
recombinant enzyme, 3 mM, approx. 75% inhibition
tyrosine
-
recombinant enzyme, 3 mM, approx. 50% inhibition
additional information
-
not inhibited by the nitric oxide synthase inhibitors N(G)-monomethyl-D-arginine, 7-nitroindazole, L-thiocitrulline, N(G)-monomethyl-L-arginine, (S)-methy-L-thiocitrulline, and L-N5-(1-iminoethyl)ornithine; PH8, the large aromatic amino acid hydroxylase monoclonal antibody, inhibits the production of both L-tyrosine and S-carboxymethyl-L-cysteine S-oxide to 99.0% and 99.3% at 0.2 mg/ml concentration
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cAMP-dependent protein kinase
-
glycerol
-
does no affect the wild-type enzyme activity at 1-5%, but increases the activity of the mutant enzymes about 1-3fold, overview
H2O2
-
2 mM, up to 4fold increase in activity, mixed activation mechanism, oxidation of Trp120 to 5-hydroxy-Trp120
L-phenylalanine
-
-
lysolecithin
phenylalanine
Phospholipids
-
activate
tetrahydrobiopterin
-
excessive dosages of BH4 inhibit PAH under normal phenylalanine conditions in vivo and activate PAH under conditions of high phenylalanine, overview
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.003 - 0.025
(6R)-5,6,7,8-tetrahydrobiopterin
0.008 - 0.053
(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin
0.2
(7R)-5,6,7,8-tetrahydrobiopterin
-
recombinant enzyme
8.3
(S)-carboxymethyl-L-cysteine
-
wild type enzyme, in 50 mM potassium phosphate buffer, pH 6.8, at 37°C
20.3
(S)-methyl-L-cysteine
-
wild type enzyme, in 50 mM potassium phosphate buffer, pH 6.8, at 37°C
1
4-Fluorophenylalanine
-
approx. value
0.022 - 0.155
5,6,7,8-tetrahydrobiopterin
0.033
6,7-dimethyltetrahydropterin
-
-
0.01 - 0.1
6-methyltetrahydropterin
0.22
7(R,S)-tetrahydrobiopterin
-
pH 7.0, 25°C, recombinant enzyme
0.008 - 0.028
Abz-VAA
3.1
L-methionine
-
at 37°C, 50 mM potassium phosphate buffer, pH 6.8
0.1 - 3
L-Phe
0.022 - 7.14
L-phenylalanine
29.8
N-acetyl-(S)-carboxymethyl-L-cysteine
-
wild type enzyme, in 50 mM potassium phosphate buffer, pH 6.8, at 37°C
32.1
N-acetyl-(S)-methyl-L-cysteine
-
wild type enzyme, in 50 mM potassium phosphate buffer, pH 6.8, at 37°C
57.15 - 63.8
N-acetyl-S-carboxymethyl-L-cysteine
60.54 - 68.25
N-acetyl-S-methyl-L-cysteine
0.043 - 0.55
phenylalanine
4.6
S-carboxy-methyl-L-cysteine
-
at 37°C, 50 mM potassium phosphate buffer, pH 6.8
0.0728 - 25.24
S-carboxymethyl-L-cysteine
0.3
S-methyl-ergothionine
-
at 37°C, 50 mM potassium phosphate buffer, pH 6.8
18.32 - 51.6
S-methyl-L-cysteine
0.0026 - 0.082
tetrahydrobiopterin
0.024 - 0.096
tryptophan
additional information
L-phenylalanine
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0183 - 6.58
phenylalanine
additional information
additional information
-
wild-type PAH kinetic analyses using a new assay reveal cooperativity of activated PAH toward (6R)-L-erythro-5,6,7,8-tetrahydrobiopterin
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0015
(7R)-5,6,7,8-tetrahydrobiopterin
-
-
1.1
4-Chlorophenylalanine
-
-
0.0023 - 0.0049
7(S)-tetrahydrobiopterin
-
pH 7.0, 25°C, recombinant enzyme, versus (6R)-tetrahydrobiopterin
3.5
L-methionine
-
at 37°C, 50 mM potassium phosphate buffer, pH 6.8
5.01
S-carboxy-methyl-L-cysteine
-
at 37°C, 50 mM potassium phosphate buffer, pH 6.8
17.23
S-carboxymethyl-L-cysteine
-
in pooled hepatic cytosolic enzyme fraction, at 37°C
0.41
S-methyl-ergothionine
-
at 37°C, 50 mM potassium phosphate buffer, pH 6.8
17.32
S-methyl-L-cysteine
-
at 37°C, 50 mM potassium phosphate buffer, pH 6.8
additional information
additional information
-
inhibition kinetics
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0009
-
mutant enzyme V388M, using S-carboxymethyl-L-cysteine as substrate
0.0014
-
mutant enzyme I65T, using S-carboxymethyl-L-cysteine as substrate; mutant enzyme R68S, using S-carboxymethyl-L-cysteine as substrate
0.0015
-
mutant enzyme Y414C, using S-carboxymethyl-L-cysteine as substrate
0.0016
-
mutant enzyme R261Q, using S-carboxymethyl-L-cysteine as substrate
0.0049
-
L-phenylalanine-activated mutant enzyme S231F, at 25°C
0.0052
-
non-L-phenylalanine-activated mutant enzyme S231F, at 25°C
0.055
-
I65T mutant enzyme, cofactor 6-methyltetrahydropterin
0.073
-
wild type enzyme, using S-carboxymethyl-L-cysteine as substrate
0.099
-
R270K mutant enzyme, expression in the absence of glycerol in the growth medium, cofactor 6-methyltetrahydropterin
0.106
-
-
0.225
-
liver enzyme
0.23
-
R270K mutant enzyme, expression in the presence of glycerol in the growth medium, cofactor 6-methyltetrahydropterin
0.408
-
V388M mutant enzyme, cofactor tetrahydrobiopterin
0.4169
-
non-L-phenylalanine-activated wild type enzyme, at 25°C
0.424
-
maltose-binding-protein phenylalanine hydroxylase fusion protein, dimeric form
0.505
-
mutant enzyme V388M, using L-phenylalanine as substrate
0.536
-
R261Q mutant enzyme, cofactor tetrahydrobiopterin
0.7418
-
L-phenylalanine-activated wild type enzyme, at 25°C
0.745
-
V388M mutant enzyme, cofactor 6-methyltetrahydropterin
0.75
-
mutant enzyme R408W, using L-phenylalanine as substrate, in 100 mM Na-HEPES buffer, pH 7.0, at 25°C
0.77
-
mutant R68V, preincubation with L-phenylalanine
0.78
-
R261Q mutant enzyme, cofactor 6-methyltetrahydropterin
1.08
-
mutant C237R
1.1
-
wild-type, preincubation with L-phenylalanine
1.13
-
mutant C237R, preincubation with L-phenylalanine
1.2
-
mutant enzyme Y414C, using L-phenylalanine as substrate
1.283
-
maltose-binding-protein phenylalanine hydroxylase fusion protein, tetrameric form
1.32
-
mutant enzyme R155H, using L-phenylalanine as substrate, in 100 mM Na-HEPES buffer, pH 7.0, at 25°C
1.49
-
mutant enzyme R261Q, using L-phenylalanine as substrate
1.6
-
fetal liver enzyme
1.64
-
mutant enzyme D143G, using L-phenylalanine as substrate, in 100 mM Na-HEPES buffer, pH 7.0, at 25°C
1.7
-
pH 7.0, 25°C, wild-type, dimer
1.725
-
mutant enzyme R68S, using L-phenylalanine as substrate
1.742
-
recombinant wild-type enzyme, cofactor tetrahydrobiopterin
1.76
-
adult liver enzyme
1.773
-
V388M mutant enzyme, expression in the absence of glycerol in the growth medium, cofactor 6-methyltetrahydropterin
1.9
-
wild type enzyme, using L-phenylalanine as substrate
1.98
-
substrate L-phenylalanine, mutant N223D
2.14
-
mutant R68V
2.2
-
mutant R68A, preincubation with L-phenylalanine
2.25
-
mutant enzyme I65T, using L-phenylalanine as substrate
2.34
-
substrate tetrahydrobiopterin, mutant N223D
2.49
-
recombinant wild-type enzyme, cofactor 6-methyltetrahydropterin
2.6
-
recombinant enzyme
2.76
-
mutant enzyme L348V, using L-phenylalanine as substrate, in 100 mM Na-HEPES buffer, pH 7.0, at 25°C
2.84
-
substrate tetrahydrobiopterin, mutant T427P
2.91
-
V388M mutant enzyme, expression in the presence of glycerol in the growth medium, cofactor 6-methyltetrahydropterin
2.94
-
mutant R68A
2.97
-
mutant C237A
2.98
-
wild type enzyme, using L-phenylalanine as substrate, in 100 mM Na-HEPES buffer, pH 7.0, at 25°C
3.17
-
substrate L-phenylalanine, mutant T427P
3.25
-
wild-type
3.32
-
mutant enzyme P416Q, using L-phenylalanine as substrate, in 100 mM Na-HEPES buffer, pH 7.0, at 25°C
3.48
-
substrate L-phenylalanine, wild-type
3.67
-
substrate L-phenylalanine, mutant N426D
3.74
-
substrate L-phenylalanine, mutant N32D
3.81
-
mutant C237D
4.19
-
mutant C237D, preincubation with L-phenylalanine
4.37
-
substrate L-phenylalanine, mutant G33A
4.39
-
substrate tetrahydrobiopterin, mutant N426D
4.73
-
substrate tetrahydrobiopterin, mutant G33A; substrate tetrahydrobiopterin, mutant N32D
4.95
-
pH 7.0, 25°C, wild-type, tetramer
5.16
-
substrate L-phenylalanine, mutant G33V
5.32
-
substrate tetrahydrobiopterin, wild-type
6.19
-
substrate L-phenylalanine, mutant K113P
6.41
-
substrate tetrahydrobiopterin, mutant G33V
6.48
-
pH 7.0, 25°C, N-terminal deletion mutant
8.32
-
pH 7.0, 25°C, N-terminal plus C-terminal deletion mutant
8.56
-
substrate tetrahydrobiopterin, mutant K113P
14
-
truncated enzyme containing C-terminal 334 amino acids
290
-
mutant Y325A, 25°C, pH 7.0
300
-
mutant Y325A, preincubation with L-phenylalanine, 25°C, pH 7.0
1150
-
mutant Y325F, 25°C, pH 7.0
1230
-
wild-type, 25°C, pH 7.0
1310
-
mutant Y325L, 25°C, pH 7.0
1500
-
mutant Y325L, preincubation with L-phenylalanine, 25°C, pH 7.0
3630
-
mutant Y325F, preincubation with L-phenylalanine, 25°C, pH 7.0
3640
-
wild-type, preincubation with L-phenylalanine, 25°C, pH 7.0
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8
-
assay at
7
-
assay at
7.3
-
assay at
7.5
-
assay at
7.8
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9.5
-
recombinant enzyme, 25% loss of activity at pH 5.5, 15% loss of activity at pH 7.0-8.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
37
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 42
-
native and recombinant enzyme
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
melanocyte
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
49000
-
x * 49000, adult liver enzyme, SDS-PAGE; x * 52000 + x * 49000, fetal liver enzyme, SDS-PAGE
53000
-
x * 53000, recombinant enzyme, SDS-PAGE
54000
-
2 * 54000, fetal liver enzyme, SDS-PAGE
94000
-
approximately 94000 Da, SDS-PAGE
95600
-
calculated from amino acid sequence
107000
-
fetal liver enzyme, sucrose density gradient centrifugation
110000
-
fetal liver enzyme, gel filtration
150000
-
adult liver enzyme, gel filtration
160000
-
fetal liver enzyme, gel filtration
165000
-
gel filtration
275000
-
-
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
-
4 * 50000
homotetramer
oligomer
-
wild-type and mutant enzymes show different oligomeric states, from dimer to hexamer, overview
tetramer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
side-chain modification
additional information
-
self-hydroxylation mechanism at Phe325, dependent on iron at the active site
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure of ternary complex of catalytic domain, Fe2+ form, with tetrahydrobiopterin and 3-(2-thienyl)-L-alanine
-
crystal structure of the catalytic domain in its catalytic active Fe2+ form and as binary complex with tetrahydrobiopterin, 1.7 and 1.5 A resolution
-
in complex with 6(R)-L-erythro-5,6,7,8-tetrahydrobiopterin and substrate analogues 3-(2-thienyl)-L-alanine or L-norleucine
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
-
V388M mutant enzyme, 50% activity after 10 min
51
-
L348V mutant enzyme, 50% activity after 10 min
59
-
recombinant wild-type enzyme, 50% activity after 10 min
additional information
-
thermal inactivation profiles of the purified wild-type enzyme, and mutants I65T, R261Q and V388M in absence or presence of 1% glycerol or trimethylamine N-oxide, overview
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
phenylalanine stabilizes
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, enzyme concentration 1 mg/ml, several months, no loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
adult and fetal liver enzyme, monoclonal antibody affinity chromatography
-
ammonium sulfate, Phenyl-Sepharose, DEAE-Sepharose, recombinant enzyme
-
amylose column chromatography
-
double truncated mutant enzyme DELTAN1-102/DELTAC428-452
-
fetal, newborn and adult enzyme are probably identical
-
HiTrap Q column chromatography
-
Phenyl-Sepharose, DEAE-Sepharose
-
recombinant enzyme, partially purified
-
recombinant His-tagged enzyme in fusion with HIV transactivator of transcription protein from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant His-tagged wild-type, I65T, R261Q and V388M mutant enzymes, affinity chromatography
-
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography
-
recombinant His6-tagged, catalytically active, heterozygous enzymes with one mutated allele, from bacteria by nickel affinity chromatography, excision of the His-tag
-
recombinant maltose-binding-protein phenylalanine hydroxylase fusion protein
-
recombinant wild-type enzyme and deletion mutants
-
recombinant wild-type, L348V and V388M mutant enzyme, affinity chromatography
-
recombinant wild-type, R270K and V388M mutant enzymes expressed in the presence and absence of glycerol
-
Superdex 200 gel filtration
-
truncated enzyme containing the C-terminal 343 amino acids
-
using Ni-NTA chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence analysis of wild-type and mutant enzymes, expression of His6-tagged, catalytically active, heterozygous enzymes with one mutated allele
-
DNA and amino acid sequence determination and analysis of wild-type and mutant enzymes, overview
-
DNA and amino acid sequence determination and analysis, genotyping, expression of mutant enzymes in HEK-193 cells
-
expressed in Escherichia coli
-
expressed in Escherichia coli as a His-tagged fusion protein
-
expressed in Escherichia coli as a recombinant protein
-
expressed in Escherichia coli as fusion proteins with maltose-binding protein
-
expressed in Escherichia coli TB1 cells
-
expression as maltose-binding-protein fusion protein in Escherichia coli circumvents proteolytic degradation by the host cell
-
expression in Escherichia coli
-
expression of double truncated mutant enzyme DELTAN1-102/DELTAC428-452 in Escherichia coli
-
expression of His-tagged wild-type, I65T, R261Q and V388M mutant enzymes in Escherichia coli
-
expression of wild-type and T427P mutant enzyme in Escherichia coli
-
expression of wild-type enzyme and Asp112-Lys452, Ser2-Gln428 and Gly103-Gln428 deletion mutants in Escherichia coli
-
expression of wild-type, L348V, L349L and V388M mutant enzyme maltose-binding-protein fusions in Escherichia coli and COS cells
-
expression of wild-type, R270K and V388M mutant enzymes in the presence of the chemical chaperone glycerol
-
gene pah, DNA and amino acid sequence determination and analysis, genotyping of a Southern Italian population, overview
-
gene pah, DNA and amino acid sequence determination and analysis, genotyping, overview
-
gene pah, DNA and amino acid sequence determination of wild-type and mutant enzymes, genotyping
-
gene pah, expression of His-tagged enzyme in fusion with HIV transactivator of transcription protein in Escherichia coli strain BL21(DE3), subcloning in Escherichi acoli strain DH5alpha
-
gene pah, expression of His6-tagged wild-type and mutant enzymes in Escherichia coli
-
gene PHA, DNA sequence determination, structural and functional analyses of mutations of the PHA gene, expression of wild-type and mutant enzymes in COS-7 cells, overview
-
identification and sequence analysis of enzyme mutant genes with exon deletions isolated from 59 czech phenylketonuria patients, multiplex ligation-dependent probe amplification method
-
mutant enzymes are expressed in Escherichia coli
-
truncated enzyme containing the C-terminal 336 amino acids bearing the catalytic domain
-
wild type and mutant enzyme S231F are expressed in Escherichia coli BL21(DE3) and human hepatoma cells, in the cells supplemented with sepiapterin the amount of expressed S231F PAH increases up to 25%
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A202T
-
the mutation is associated with phenylketonuria
A259V
-
the mutant with wild type activity exhibits less than 50% of wild type protein level and leads to classic phenyletonuria
A300S
-
naturally occuring mutation involved in hyperphenylalaninemia and/or in phenylketonuria, overview
A309V
-
the mutant shows 70% of wild type activity
A322G
-
the mutant shows 75% of wild type activity
A395G
-
naturally occuring mutation involved in hyperphenylalaninemia and/or in phenylketonuria, overview
C237A
-
increase of basal activity and affinity for substrate L-phenylalanine
C237R
-
reduced activity, elimination of positive cooperativity
C237S
-
approx. 2fold higher activity than wild-type
D143G
-
mutant with a mild misfolding defect associated with phenylketonuria
D415N
-
naturally occuring missense mutation causing a mild phenylketonuria phenotype
DELTA1-102
-
mutant lacking the first 102 residues corresponding to the N-terminal regulatory domain. 96% of the truncated mutant exist as a tetramer. On coexpression of wild-type-hPAH and the N-terminally truncated form DELTA1-102 (~95% tetramer), heterotetramers, as a result of an assembly of two different homodimers, are isolated. The recovered (wild-type)/(DELTA1-102 mutant)-hPAH heterotetramers reveal a catalytic activity deviating significantly from that calculated by averaging the respective recombinant homotetrameric forms. The heterotetramer assembly also results in conformational changes in the WT-hPAH protomer, as detected by trypsin limited proteolysis
DELTA1-102/DELTAC24
-
mutant lacking the first 102 residues corresponding to the N-terminal regulatory domain and the last 24 residues at the C-terminal end corresponding to the tetramerisation motif. 81% of the truncated mutant exist as a dimer and 17% as an aggregated form. On co-expression of wild-type-hPAH (50% tetramer, 10% dimer) and the N- and C-terminally truncated form DELTA1-102/DELTAC24 (80% dimer) no heterodimers is recovered
DELTA103-427
-
dimeric double-truncated form: the dimeric variant 103-427 shows a Vmax (1980 nmol Tyr/min/mg protein) comparable with that of the non-activated wild-type PAH, which does not change markedly upon L-Phe preincubation (2421 nmol Tyr/min/mg protein)
E178G
-
exon 6 A533G mutation naturally occuring in phenylketonuria patients from the Cukurova region in Turkey, sequence determination and analysis
E280K
-
inactive; the mutant shows 2% of wild type activity
E76G
-
the mutant shows 65% of wild type activity; the mutant shows 85% of wild type activity
F161S
-
the mutant with wild type activity exhibits less than 50% of wild type protein level
F382L
-
naturally occuring mutation and site-directed mutagenesis, the mutant shows 44% reduced activity compared to the wild-type enzyme, analysis of structural alterations
F39C
-
the mutant enzyme shows reduced activity compared to the wild type
F39L/F55fsdelT
-
naturally occuring mutation in the regulatory domain, that affects enzyme activity and causes an atypical form of phenylketonuria
F39L/P281L
-
naturally occuring mutation in the regulatory domain, that affects enzyme activity and causes the classical form of phenylketonuria
F39L/R408W
-
naturally occuring mutation in the regulatory domain, that affects enzyme activity and causes the classical form of phenylketonuria
G103S
-
site-directed mutagenesis, the mutation occurs naturally in phenylketonuria patients from Korea, the mutant shows highly reduced activity compared to the wild-type
G218V
-
the mutant shows wild type activity
G247R
-
the mutation is associated with phenylketonuria
G247V
-
the mutant shows 4% of wild type activity
G332E
-
the mutation is associated with phenylketonuria
G332V
-
site-directed mutagenesis, the mutation occurs naturally in phenylketonuria patients from Korea, inactive mutant
G33A
-
increased basal activity, reduced activation by preincubation with substrate
G33V
-
increased basal activity, reduced activation by preincubation with substrate
G344D
-
the mutation is associated with phenylketonuria
H271Q
-
naturally occuring knockout missense mutation leading to a severe phenylketonuria phenotype
I174V
-
naturally occuring missense mutation causing a mild phenylketonuria phenotype
I65S
-
the mutant enzyme shows reduced activity compared to the wild type
I65T/R408W
I65T/R68S
-
naturally occuring mutation in the regulatory domain, that affects enzyme activity and causes a mild form of phenylketonuria
I65V
-
the mutant enzyme shows reduced activity compared to the wild type
I95F
-
naturally occuring missense mutation causing a mild phenylketonuria phenotype
I97L
-
naturally occuring mutation in the regulatory domain, that affects enzyme activity and is involved in the disorder hyperphenylalaninemia
K113P
-
increased basal activity, reduced activation by preincubation with substrate, increase in positive cooperativity
K398K
-
naturally occuring mutation
K398N
-
naturally occuring mutation and site-directed mutagenesis, the mutant shows 45% reduced activity compared to the wild-type enzyme, analysis of structural alterations
K42I
-
the mutant shows 12% of wild type activity
L197F
-
naturally occuring knockout missense mutation leading to a severe phenylketonuria phenotype
L212P
naturally occuring mutation involved in phenylketonuria
L255S
-
the mutant shows 3% of wild type activity
L255V
-
the mutant shows 13% of wild type activity
L293M
-
site-directed mutagenesis, the mutation occurs naturally in phenylketonuria patients from Korea, the mutant shows reduced activity and no response to tetrahydrobiopterin compared to the wild-type
L311P
-
the mutant with wild type activity exhibits less than 50% of wild type protein level and leads to classic phenyletonuria
L41F
-
the mutant shows 10% of wild type activity
N223D
-
low basal activity, little activation by preincubation with substrate, increase in positive cooperativity
N223Y
-
naturally occuring mutation and site-directed mutagenesis, the mutant shows 30% reduced activity compared to the wild-type enzyme, analysis of structural alterations
N32D
-
low basal activity, close to normal activation by preincubation with substrate
N426D
-
low basal activity, close to normal activation by preincubation with substrate
P122Q
-
the mutant with wild type activity exhibits less than 50% of wild type protein level
P225T
-
naturally occuring knockout missense mutation leading to a severe phenylketonuria phenotype
P244L
-
the mutant shows 68% of wild type activity
P366H
-
naturally occuring mutation involved in hyperphenylalaninemia and/or in phenylketonuria, overview
P416Q
-
the mutant retains significant catalytic activity yet is observed in classic and moderate phenylketonuria patients
P69S
-
site-directed mutagenesis, the mutation occurs naturally in phenylketonuria patients from Korea, the mutant shows reduced activity compared to the wild-type
Q419R
-
naturally occuring mutation and site-directed mutagenesis, the mutant shows 29% reduced activity compared to the wild-type enzyme, analysis of structural alterations
R111X
-
the mutation is associated with phenylketonuria
R155H
-
the mutant displays low PAH activity and decreased apparent affinity for L-Phe yet is observed in mild hyperphenylalaninaemia, mutant does not display kinetic instability, as it is stabilized by (6R)-L-erythro-5,6,7,8-tetrahydrobiopterin similarly to wild type enzyme
R157N
-
the mutant with wild type activity exhibits less than 50% of wild type protein level
R158W
-
naturally occuring mutation involved in hyperphenylalaninemia and/or in phenylketonuria, overview
R176X
-
the mutation is associated with phenylketonuria
R243X
-
exon 6 C727T mutation naturally occuring in phenylketonuria patients from the Cukurova region in Turkey, sequence determination and analysis
R252G
-
the mutant with wild type activity exhibits less than 50% of wild type protein level and leads to classic phenyletonuria
R252Q
-
the mutant with wild type activity exhibits less than 50% of wild type protein level and leads to classic phenyletonuria
R261P
-
naturally occuring missense mutation causing a mild phenylketonuria phenotype
R270S
-
the mutant shows 3% of wild type activity
R297C
-
naturally occuring mutation
R297H
-
naturally occuring mutation
R297L
-
naturally occuring mutation and site-directed mutagenesis, the mutant shows 58% reduced activity compared to the wild-type enzyme, analysis of structural alterations
R408W/I283F
-
the mutant shows 2% residual activity compared to the wild type enzyme
R408W/I306V
-
the mutant shows 18% residual activity compared to the wild type enzyme
R408W/P281L
-
inactive
R408W/pA403V
-
the mutant shows 20% residual activity compared to the wild type enzyme
R408W/R158Q
-
the mutant shows 5% residual activity compared to the wild type enzyme
R408W/R297H
-
the mutant shows 15% residual activity compared to the wild type enzyme
R408W/R408W
-
inactive
R53H
-
site-directed mutagenesis, the mutation occurs naturally in humans altering the tetrahydrobiopterin responsiveness, the mutant shows reduced activity and dimer stability compared to the wild-type
R68A
-
increase of basal activity and affinity for substrate L-phenylalanine
R68G
-
the mutant shows wild type activity
R68S/R408W
-
naturally occuring mutation in the regulatory domain, that affects enzyme activity and causes an atypical form of phenylketonuria
R68V
-
little decrease in activity
R71C
-
naturally occuring mutation in the regulatory domain, that affects enzyme activity and is involved in the disorder hyperphenylalaninemia
R86S
-
for the variants R68S and V106A, a Vmax comparable with the activated wild-type PAH is found without L-Phe preincubation, and no further increase is measured when the substrate is present. R68S and V106A without L-Phe preincubation show lower cofactor affinities than the non-activated wild-type PAH. Values are at the same level as determined for the L-Phe preincubated wild-type PAH
S231F
-
the missense phenylalanine hydroxylase gene mutation causes complete loss of enzymatic activity in vitro (residual enzyme activity in vitro is about 1%) as it drastically reduces stability and activity of the PAH enzyme, the mutant enzyme is not activated by pre-incubation with L-phenylalanine substrate
S348L
-
instable enzyme forming aggregates after expression in Escherichia coli in the presence of GroESL
S349L
-
inactive
S391I
-
site-directed mutagenesis, the mutation occurs naturally in phenylketonuria patients from Korea, inactive mutant
T278I
-
the mutation is associated with phenylketonuria
V106A
-
for the variants R68S and V106A, a Vmax comparable with the activated wild-type PAH is found without L-Phe preincubation, and no further increase is measured when the substrate is present. R68S and V106A without L-Phe preincubation show lower cofactor affinities than the non-activated wild-type PAH. Values are at the same level as determined for the L-Phe preincubated wild-type PAH
V379D/H264Q
-
the mutant shows significant activity at tyrosine hydroxylation and a 3000fold decrease in preference for phenylalanine over tyrosine as the substrate
W187X
-
the mutation is associated with phenylketonuria
Y166X
-
the mutation is associated with phenylketonuria
Y168H
-
the mutation is associated with phenylketonuria
Y204C
-
the mutation is associated with phenylketonuria
Y277D
-
inactive
Y325L
-
stable, similar yields and oligomeric distribution as wild-type, reduced specific activity, decreased coupling efficiency and decreased iron content, no positive cooperativity for L-phenylalanine
Y325S
-
aggregation after purification, not suitable for characterization
Y356X
-
the mutation is associated with phenylketonuria
Y386C
-
exon 11 A1157G mutation naturally occuring in phenylketonuria patient from the Cukurova region in Turkey, sequence determination and analysis
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
molecular biology