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Information on EC 4.1.99.2 - tyrosine phenol-lyase
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4.1.99.2 tyrosine phenol-lyaseIUBMB Comments
A pyridoxal-phosphate protein. The enzyme cleaves a carbon-carbon bond, releasing phenol and an unstable enamine product that tautomerizes to an imine form, which undergoes a hydrolytic deamination to form pyruvate and ammonia. The latter reaction, which can occur spontaneously, can also be catalysed by EC 3.5.99.10, 2-iminobutanoate/2-iminopropanoate deaminase. The enzyme also slowly catalyses similar reactions with D-tyrosine, S-methyl-L-cysteine, L-cysteine, L-serine and D-serine.
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Word Map
- 4.1.99.2
- triptolide
- citrobacter
-
preterm
- freundii
-
two-photon
-
topless
- tripterygium
- wilfordii
-
quinonoid
-
nitroxide
- herbicola
-
beta-elimination
-
indole-lyase
-
co-repressors
- tryptophanase
-
aldimine
-
nanorods
-
tempol
-
tocolysis
-
photoluminescence
-
triepoxide
-
phillips
-
disaturated
- 3,4-dihydroxyphenyl-l-alanine
- synthesis
- biotechnology
- degradation
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Synonyms
beta-tyrosinase, Fn-TPL, L-tyrosine phenol-lyase, phenol-lyase, tyrosine, TnaA, TPL, tyrosine phenol lyase, tyrosine phenol-lyase, tyrosine-phenol lyase, 
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
beta-tyrosinase
Fn-TPL
L-tyrosine phenol-lyase
phenol-lyase, tyrosine
-
-
-
-
TnaA
TPL
tyrosine phenol lyase
tyrosine phenol-lyase
beta-tyrosinase
-
-
-
-
L-tyrosine phenol-lyase
-
-
-
-
TnaA
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
gene name; gene name
-
TPL
-
-
-
-
tyrosine phenol lyase
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
-
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
L-tyrosine + H2O = phenol + pyruvate + NH3
-
-
-
-
L-tyrosine + H2O = phenol + pyruvate + NH3
mechanism appears to require two bases, one of which abstracts the proton from the 2-position of the substrate to form a quinonoid intermediate and the second of which acting in concert with proton transfer from the first group to C-1 of the phenolic ring, abstracts the substrate hydroxyl group to facilitate cyclohexadienone formation and subsequent elimination of phenol
-
L-tyrosine + H2O = phenol + pyruvate + NH3
mechanism appears to require two bases, one of which abstracts the proton from the 2-position of the substrate to form a quinonoid intermediate and the second of which acting in concert with proton transfer from the first group to C-1 of the phenolic ring, abstracts the substrate hydroxyl group to facilitate cyclohexadienone formation and subsequent elimination of phenol
-
L-tyrosine + H2O = phenol + pyruvate + NH3
single-base racemization mechanism, proceeds with retention of configuration at C-beta
-
L-tyrosine + H2O = phenol + pyruvate + NH3
NH3 is the first substrate which interacts with bound pyridoxal 5'-phosphate, pyruvate is the second substrate
-
L-tyrosine + H2O = phenol + pyruvate + NH3
mechanism
-
L-tyrosine + H2O = phenol + pyruvate + NH3
mechanism
-
L-tyrosine + H2O = phenol + pyruvate + NH3
reversible cleavage of the Cbeta-Cgamma bond of L-tyrosine in the final stage of the beta-elimination, via intermediates: internal aldimine, external aldimine, quinoid, ketoquinoid, alpha-aminoacrylate, and internal aldimine, stepwise mechanism, overview
-
L-tyrosine + H2O = phenol + pyruvate + NH3
mechanism of alpha,beta-elimination of L-tyrosine catalyzed by enzyme TPL, detailed overview
-
L-tyrosine + H2O = phenol + pyruvate + NH3
mechanism of alpha,beta-elimination of L-tyrosine catalyzed by enzyme TPL
-
L-tyrosine + H2O = phenol + pyruvate + NH3
an active site base is essential for activity, and alpha-deuterated substrate exhibits modest primary isotope effects on kcat and kcat/Km, suggesting that substrate deprotonation is partially rate-limiting. Pre-steady state kinetics with enzyme TPL show rapid formation of external aldimine intermediate, followed by deprotonation to give a quinonoid intermediate absorbing at about 500 nm. The mechanism of TPL requires both substrate strain and acid/base catalysis, and substrate strain is probably responsible for the very high substrate specificity of TPL. Acid-base catalysis mechanism of TPL, overview
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
alpha,beta-elimination
racemization
transamination
alpha,beta-elimination
-
-
-
-
racemization
-
-
-
-
transamination
-
-
-
-
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SYSTEMATIC NAME
IUBMB Comments
L-tyrosine phenol-lyase (deaminating; pyruvate-forming)
A pyridoxal-phosphate protein. The enzyme cleaves a carbon-carbon bond, releasing phenol and an unstable enamine product that tautomerizes to an imine form, which undergoes a hydrolytic deamination to form pyruvate and ammonia. The latter reaction, which can occur spontaneously, can also be catalysed by EC 3.5.99.10, 2-iminobutanoate/2-iminopropanoate deaminase. The enzyme also slowly catalyses similar reactions with D-tyrosine, S-methyl-L-cysteine, L-cysteine, L-serine and D-serine.
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CAS REGISTRY NUMBER
COMMENTARY
9059-31-8
-
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
1-amino-2-(4-hydroxyphenyl)-ethyl phosphinic acid + H2O
phenol + ?
-
-
-
?
2,5-difluorotyrosine + H2O
2,5-difluorophenol + pyruvate + NH3
-
-
-
?
2,6-difluorotyrosine + H2O
2,6-difluorophenol + pyruvate + NH3
-
-
-
?
2-fluorotyrosine + H2O
2-fluorophenol + pyruvate + NH3
-
-
-
?
3,4-dihydroxyphenyl-L-alanine + H2O
pyrocatechol + NH3 + pyruvate
-
-
-
-
?
3,5-difluorotyrosine + H2O
3,5-difluorophenol + pyruvate + NH3
-
-
-
?
3-chloro-L-tyrosine + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
3-chlorotyrosine + H2O
3-chlorophenol + pyruvate + NH3
-
-
-
?
3-fluoro-L-tyrosine + H2O
3-fluoro-phenol + pyruvate + NH3
-
-
?
3-fluoro-L-tyrosine + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
3-fluorotyrosine + H2O
3-fluorophenol + pyruvate + NH3
-
-
-
?
3-methoxy-L-tyrosine + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
3-methyl-L-tyrosine + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
L-Ala + pyridoxal phosphate
pyridoxamine phosphate + keto acid
-
-
-
-
ir
L-alanine + H2O
?
-
wild-type enzyme forms the Ala quinonoid intermediate when incubated with L-Ala
-
-
?
L-cysteine + H2O
thiophenol + pyruvate + NH3
-
12% of the activity with L-tyrosine
-
-
r
L-DOPA + H2O
pyrocatechol + pyruvate + NH4+
-
42% of the activity with L-tyrosine
-
-
r
L-m-Tyr + pyridoxal phosphate
pyridoxamine phosphate + keto acid
-
-
-
-
ir
L-Phe + pyridoxal phosphate
pyridoxamine phosphate + keto acid
-
-
-
-
ir
L-phenylalanine + H2O
? + pyruvate + NH3
-
7% of the activity with L-tyrosine
-
-
r
L-Ser + pyridoxal phosphate
pyridoxamine phosphate + keto acid
-
-
-
-
ir
L-serine + H2O
pyruvate + NH3
-
9% of the activity with L-tyrosine
-
-
r
L-valine + H2O
? + pyruvate + NH3
-
6% of the activity with L-tyrosine
-
-
r
o-chlorophenol + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
o-cresol + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
o-methoxyphenol + H2O
?
-
substrate of mutant M379V, not of the wild-type enzyme
-
-
?
S-methyl-L-cysteine + H2O
?
Symbiobacterium sp.
-
385% of activity compared to L-tyrosine
-
?
S-methyl-L-cysteine + H2O
methanthiol + pyruvate + NH3
-
13% of the activity with L-tyrosine
-
-
r
3,4-dihydroxyphenyl-L-Ala + H2O
pyrocatechol + NH3 + pyruvate
-
-
-
?
3,4-dihydroxyphenyl-L-Ala + H2O
pyrocatechol + NH3 + pyruvate
-
i.e. L-dopa
-
?
3,4-dihydroxyphenyl-L-Ala + H2O
pyrocatechol + NH3 + pyruvate
-
i.e. L-dopa
-
?
3-fluoro-L-tyrosine + H2O
3-fluorophenol + pyruvate + NH3
-
-
-
-
?
3-fluoro-L-tyrosine + H2O
3-fluorophenol + pyruvate + NH3
-
-
-
-
r
3-fluoro-L-tyrosine + H2O
3-fluorophenol + pyruvate + NH3
-
3-fluoro-L-tyrosine also is a good substrate for the wild-type enzyme
-
-
r
beta-chloro-L-alanine + H2O
?
Symbiobacterium sp.
-
500% of activity compared to L-tyrosine
-
?
beta-chloro-L-alanine + H2O
?
-
500% of activity compared to L-tyrosine
-
?
catechol + pyruvate + NH3
3,4-dihydroxyphenyl-L-alanine
-
TPL is well-correlated to cytoplasmic 3,4-dihydroxyphenyl-L-alanine levels
-
-
r
catechol + pyruvate + NH3
3,4-dihydroxyphenyl-L-alanine
-
TPL is well-correlated to cytoplasmic 3,4-dihydroxyphenyl-L-alanine levels
-
-
r
D-tyrosine + H2O
phenol + pyruvate + NH3
Symbiobacterium sp.
-
at 7% the rate of L-tyrosine
-
?
D-tyrosine + H2O
phenol + pyruvate + NH3
-
at 7% the rate of L-tyrosine
-
?
L-2-aminoadipate + H2O
?
-
no activity with wild-type enzyme, activity with mutant enzyme R100T/V283R
-
-
?
L-2-aminoadipate + H2O
?
-
no activity with wild-type enzyme, activity with mutant enzyme R100T/V283R
-
-
?
L-3-4 dihydroxyphenylalanine + H2O
catechol + pyruvate + NH3
-
L-dopa
-
-
r
L-3-4 dihydroxyphenylalanine + H2O
catechol + pyruvate + NH3
-
L-DOPA, alpha-beta elimination
-
-
r
L-3-4 dihydroxyphenylalanine + H2O
catechol + pyruvate + NH3
-
L-dopa
-
-
r
L-Asp + H2O
formate + pyruvate + NH3
-
no activity with wild-type enzyme, activity with mutant enzyme R100T/V283R
-
?
L-Asp + H2O
formate + pyruvate + NH3
-
no activity with wild-type enzyme, activity with mutant enzyme R100T/V283R
-
?
L-Glu + H2O
?
-
no activity with wild-type enzyme, activity with mutant enzyme R100T/V283R
-
-
?
L-Glu + H2O
?
-
no activity with wild-type enzyme, activity with mutant enzyme R100T/V283R
-
-
?
L-Ser + phenol
L-Tyr + H2O
-
-
-
?
L-Ser + pyrocatechol
3,4-dihydroxyphenyl-L-Ala + H2O
-
-
i.e. L-dopa
?
L-Ser + pyrocatechol
3,4-dihydroxyphenyl-L-Ala + H2O
-
-
-
?
L-tryptophan + H2O
indole + pyruvate + NH3
-
8% of the activity with L-tyrosine, reaction of EC 4.1.99.1
-
-
r
L-tryptophan + H2O
indole + pyruvate + NH3
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
-
8% of the activity with L-tyrosine, reaction of EC 4.1.99.1
-
-
r
L-tyrosine + H2O
3,4-dihydroxyphenyl-L-alanine
-
TPL is well-correlated to cytoplasmic 3,4-dihydroxyphenyl-L-alanine levels
-
-
?
L-tyrosine + H2O
3,4-dihydroxyphenyl-L-alanine
-
TPL is well-correlated to cytoplasmic 3,4-dihydroxyphenyl-L-alanine levels
-
-
?
L-tyrosine + H2O
phenol + pyruvate + NH3
-
-
-
-
?
L-tyrosine + H2O
phenol + pyruvate + NH3
-
-
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
-
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
alpha-beta elimination
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
reversible cleavage of the Cbeta-Cgamma bond of L-tyrosine in the final stage of the beta-elimination, via intermediates: internal aldimine, external aldimine, quinoid, ketoquinoid, alpha-aminoacrylate, and internal aldimine, overview
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
very high substrate specificity of TPL
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
high activity with the wild-type enzyme
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
alpha-beta elimination
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
L-tyrosine is the physiological substrate of enzyme Fn-TPL
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
alpha,beta-elimination of L-tyrosine, best substrate
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
-
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
-
L-tyrosine is the physiological substrate of enzyme Fn-TPL
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
-
alpha,beta-elimination of L-tyrosine, best substrate
-
-
r
pyrocatechol + pyruvate + NH3
3,4-dihydroxyphenyl-L-Ala + H2O
-
-
-
?
pyrocatechol + pyruvate + NH3
3,4-dihydroxyphenyl-L-Ala + H2O
-
-
-
?
pyrocatechol + pyruvate + NH3
3,4-dihydroxyphenyl-L-Ala + H2O
-
-
-
?
pyrocatechol + pyruvate + NH4+
L-DOPA + H2O
-
-
-
r
pyrocatechol + pyruvate + NH4+
L-DOPA + H2O
-
-
-
-
r
pyrocatechol + pyruvate + NH4+
L-DOPA + H2O
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
-
-
-
r
pyrocatechol + pyruvate + NH4+
L-DOPA + H2O
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
-
-
-
-
r
S-(2-nitrophenyl)-L-cysteine + H2O
2-nitrobenzenethiolate + pyruvate + NH3
-
-
-
-
r
S-(2-nitrophenyl)-L-cysteine + H2O
2-nitrobenzenethiolate + pyruvate + NH3
-
best substrate for the wild-type enzyme
-
-
r
S-(o-nitrophenyl)-L-cysteine + H2O
?
Symbiobacterium sp.
-
250% of activity compared to L-tyrosine
-
?
S-(o-nitrophenyl)-L-cysteine + H2O
?
-
250% of activity compared to L-tyrosine
-
?
Tyr + H2O
phenol + pyruvate + NH3
-
L-Tyr
-
-
-
additional information
?
-
-
mechanism of alpha-proton isotope exchange in amino acids catalysed by tyrosine phenol-lyase
-
-
-
additional information
?
-
-
alanine, arginine, aspartate, cysteine, glutamine, glutamate, glycine, histidine, isoleucine, leucine, lysine, proline, serine, threonine, tryptophan are poor or no substrates, substrate specificity of purified tyrosine phenol lyase, overview
-
-
-
additional information
?
-
-
substrate 3-fluoro-L-tyrosine is converted only to the quinoid reaction intermediate, structure determination of 3-fluoro-L-tyrosine bound to enzyme mutant Y71F with open active site and mutant F448H with closed active site, overview
-
-
-
additional information
?
-
-
substrate specificity with 3-substituted tyrosines, the wild-type enzyme shows a narrow substrate spectrum, overview. Mutant M379V converts several L-tyrosine derivatives to the correspondent products
-
-
-
additional information
?
-
-
fragment of active site structure of the quinonoid complex of wild-type enzyme TPL with L-alanine, overview
-
-
-
additional information
?
-
-
of the common S-alkyl-L-cysteine substrates, S-ethyl-L-cysteine has the most favorable kinetic properties with TPL
-
-
-
additional information
?
-
-
tyrosine phenol-lyase is a pyridoxal 5'-phosphate (PLP)-dependent bacterial enzyme that catalyzes the reversible hydrolytic cleavage of the Cbeta-Cgamma bond of L-Tyr to phenol and ammonium pyruvate. In addition to its physiological substrate L-Tyr, TPL also catalyzes the in vitro beta-elimination of substrates with good leaving groups on the beta-carbon, including L-serine, L-cysteine, S-(2-nitrophenyl)-L-cysteine, O-acetyl-L-serines, and S-alkyl-L-cysteines
-
-
-
additional information
?
-
-
adaptive enzyme responsible for cells' growth on media with L-Tyr as the sole source of carbon
-
-
-
additional information
?
-
-
low activity of Fn-TPL is observed toward natural amino acids including L-serine, L-cysteine, L-phenylalanine, L-valine, L-tryptophan, no activity with L-threonine, L-glutamine, L-alanine, D-alanine, L-arginine, L-leucine, L-histidine, L-aspartic acid, L-methionine, and D-methionine
-
-
-
additional information
?
-
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
-
low activity of Fn-TPL is observed toward natural amino acids including L-serine, L-cysteine, L-phenylalanine, L-valine, L-tryptophan, no activity with L-threonine, L-glutamine, L-alanine, D-alanine, L-arginine, L-leucine, L-histidine, L-aspartic acid, L-methionine, and D-methionine
-
-
-
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
L-3-4 dihydroxyphenylalanine + H2O
catechol + pyruvate + NH3
-
L-dopa
-
-
r
L-3-4 dihydroxyphenylalanine + H2O
catechol + pyruvate + NH3
-
L-DOPA, alpha-beta elimination
-
-
r
L-3-4 dihydroxyphenylalanine + H2O
catechol + pyruvate + NH3
-
L-dopa
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
-
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
alpha-beta elimination
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
very high substrate specificity of TPL
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
alpha-beta elimination
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
Q8RHM6
-
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
-
L-tyrosine is the physiological substrate of enzyme Fn-TPL
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
Q8RHM6
-
-
-
r
L-tyrosine + H2O
phenol + pyruvate + NH3
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
-
L-tyrosine is the physiological substrate of enzyme Fn-TPL
-
-
r
additional information
?
-
-
adaptive enzyme responsible for cells' growth on media with L-Tyr as the sole source of carbon
-
-
-
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
pyridoxal 5'-phosphate
-
2 pyridoxal 5'-phosphate binding sites per mol of enzyme; cofactor
pyridoxal 5'-phosphate
-
2 pyridoxal 5'-phosphate binding sites per mol of enzyme; cofactor
pyridoxal 5'-phosphate
-
contains 2 mol of pyridoxal phosphate per mol of enzyme; Km: 0.00152 mM
pyridoxal 5'-phosphate
-
the phosphate group changes from being monoanionic at low pH to dianionic at high pH
pyridoxal 5'-phosphate
-
dependent on, highest activity at 0.015-0.02 mM
pyridoxal 5'-phosphate
-
dependent on, the active site residues involved in cofactor binding are highly conserved for enzyme TPL. Pyridoxal 5'-phosphate is bound to the internal aldimine by a lysine epsilon-amino group of Lys257 and the protonated pyridine ring forms an ionic/hydrogen bond with an aspartate beta-carboxylate of Asp214
pyridoxal 5'-phosphate
-
dependent on
pyridoxal 5'-phosphate
-
dependent on, activates, Km value is 0.0034 mM
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K+
NH4+
additional information
additional information
-
divalent cations (Mg2+, Ca2+, Ba2+, and Sr2+) do not activate TPL
additional information
-
the enzyme requires a monovalent cation, either K+, NH4+, Rb+ or Cs+ for activity, with Na+ and Li+ giving little or no activity. The cation is bound by the gamma-carboxylate of a conserved glutamate residue Glu69 of the TPL peptide backbone carbonyl oxygens, and waters, and is located about 10 A from cofactor pyridoxal 5'-phosphate
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3,4-dihydroxyphenyl-L-alanine
-
inactivated by a Pictet-Spengler reaction between the cofactor and 3,4-dihydroxyphenyl-L-alanine, on treatment with excess pyridoxal-5'-phosphate the inactivated enzymes recovers over 80% of the original activity
4-hydroxyphenylpropionate
-
competitive inhibitor, addition of 4-hydroxypyridin in the reaction results in decay of the quinonoid-aldimine spectrum with rate constant of 1.4/s and formation of an intermediate absorbing at 338 nm, assigned to an aminoacrylate intermediate
phenol
-
In presence of 1 mM, only 23% of the original activity remained. The presence of phenol seriously affects its own production, through the inhibition of TPL
additional information
-
inhibition of tyrosine phenol-lyase by tyrosine homologues and their O-methyl derivatives, overview. Synthesis, and steady-state and pre-steady-state kinetic evaluations of L-homotyrosine and L-bishomotyrosine as possible mechanism-based inhibitors for TPL
-
additional information
-
The mutant S51A also shows intermediate formation with L-Met and L-Phe. Influence of replacement of Ser51 by Ala on the kinetic parameters of TPL reactions with L-phenylalanine and L-methionine, kinetics and structures, overview
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
Vitreoscilla hemoglobin positive strains (vgb+ strains) have substantially higher TPL activity than the respective wild-type hosts or the strains bearing a comparable plasmid without vgb (the vgb- strains)
-
additional information
-
Vitreoscilla hemoglobin positive strains (vgb+ strains) have substantially higher TPL activity than the respective wild-type hosts or the strains bearing a comparable plasmid without vgb (the vgb- strains)
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
15.62
pyrocatechol
-
recombinant enzyme, L-DOPA formation is measured from dicatechol, pyruvate, and ammonium, pH 8.5, 20°C
3.2
3,4-dihydroxyphenyl-L-alanine
-
wild type enzyme, in 0.1 M potassium phosphate buffer (pH 8.0) at 30°C
2
3-chloro-L-Ala
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
11.4
3-chloro-L-Ala
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
13.3
3-chloro-L-Ala
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
0.1
3-fluoro-L-Tyr
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
0.2
L-Tyr
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
0.22
L-tyrosine
-
mutant enzyme T15A, in 0.1 M potassium phosphate buffer (pH 8.0) at 30°C
0.24
L-tyrosine
-
wild type enzyme, in 0.1 M potassium phosphate buffer (pH 8.0) at 30°C
3.6
phenol
-
recombinant enzyme, L-tyrosine formation is measured from phenol, pyruvate, and ammonium, pH 8.5, 20°C
0.21
S-(2-nitrophenyl)-L-cysteine
-
recombinant wild-type enzyme, pH 8.0, 30°C
2.2
S-(2-nitrophenyl)-L-cysteine
-
recombinant mutant S51A, pH 8.0, 30°C
0.1
S-(o-nitrophenyl)-L-Cys
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
1.32
S-(o-nitrophenyl)-L-Cys
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
1.86
S-(o-nitrophenyl)-L-Cys
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
0.2
S-benzyl-L-Cys
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
5.1
S-benzyl-L-Cys
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
7
S-benzyl-L-Cys
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
3.4
S-methyl-L-Cys
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
13.4
S-methyl-L-Cys
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
50
S-methyl-L-Cys
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
additional information
additional information
-
pH-independent kinetic constants for substrates bearing small substituents
-
additional information
additional information
-
pre-steady-state kinetic and steady-state kinetics, kinetic analysis of recombinant wild-type and mutant enzymes, stopped-flow kinetics of enzyme mutant F448A
-
additional information
additional information
-
pre-steady state kinetics and steady state kinetic study, overview. TPL from Citrobacter freundii exhibits two basic pKas, with an average value of 7.8, in the pH dependence of kcat/Km, and kcat is pH-independent. The TPL reaction shows primary isotope effects of about 3 on kcat and 2 on kcat/Km for both enzymes, also suggesting that steps involving transfer of the alpha-proton are partially rate-limiting. The primary isotope effect increases to 5.4 and 3.8, respectively, for kcat and kcat/Km with alpha-[2H]-3-fluoro-L-tyrosine with H343A TPL, compared to 3.9 and 2.2 for wild-type TPL. Stopped-flow measurements, the reaction of TPL with L-tyrosine and 3-fluoro-L-tyrosine in the stopped-flow spectrophotometer also shows rapid formation of quinonoid intermediate
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.44
phenol
-
recombinant enzyme, L-tyrosine formation is measured from phenol, pyruvate, and ammonium, pH 8.5, 20°C
2.68
pyrocatechol
-
recombinant enzyme, L-DOPA formation is measured from dicatechol, pyruvate, and ammonium, pH 8.5, 20°C
0.31
3,4-dihydroxyphenyl-L-alanine
-
wild type enzyme, in 0.1 M potassium phosphate buffer (pH 8.0) at 30°C
0.0065
3-chloro-L-Ala
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
0.02
3-chloro-L-Ala
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
0.7
3-chloro-L-Ala
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
1.4
3-fluoro-L-Tyr
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
3.5
L-Tyr
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
1.2
L-tyrosine
-
mutant enzyme T15A, in 0.1 M potassium phosphate buffer (pH 8.0) at 30°C
1.8
L-tyrosine
-
wild type enzyme, in 0.1 M potassium phosphate buffer (pH 8.0) at 30°C
0.26
S-(2-nitrophenyl)-L-cysteine
-
pH 8.0, 25°C, recombinant mutant F448L
0.36
S-(2-nitrophenyl)-L-cysteine
-
pH 8.0, 25°C, recombinant mutant F449A
0.6
S-(2-nitrophenyl)-L-cysteine
-
pH 8.0, 25°C, recombinant mutant F448H
2.5
S-(2-nitrophenyl)-L-cysteine
-
pH 8.0, 25°C, recombinant mutant F448A
8.7
S-(2-nitrophenyl)-L-cysteine
-
recombinant mutant S51A, pH 8.0, 30°C
9.65
S-(2-nitrophenyl)-L-cysteine
-
recombinant wild-type enzyme, pH 8.0, 30°C
9.7
S-(2-nitrophenyl)-L-cysteine
-
pH 8.0, 25°C, recombinant wild-type enzyme
0.024
S-(o-nitrophenyl)-L-Cys
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
0.041
S-(o-nitrophenyl)-L-Cys
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
5.1
S-(o-nitrophenyl)-L-Cys
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
7.4
S-(o-nitrophenyl)-L-cysteine
-
pH 8.0, 25°C, mutant enzyme E69D/K256R
0.0067
S-benzyl-L-Cys
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
0.02
S-benzyl-L-Cys
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
0.5
S-benzyl-L-Cys
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
0.0016
S-methyl-L-Cys
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
0.0052
S-methyl-L-Cys
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
0.9
S-methyl-L-Cys
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C
additional information
additional information
-
pH-independent kinetic constants for substrates bearing small substituents
-
additional information
additional information
the turnover-numbers of the mutant enzyme R381I, R381V and R381A for the substrates S-(o-nitrophenyl)-L-Cys, 3-chloro-L-Ala, O-benzoyl-L-Ser and S-methyl-L-Cys are comparable to those of the wild-type enzyme
-
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.12
phenol
-
recombinant enzyme, L-tyrosine formation is measured from phenol, pyruvate, and ammonium, pH 8.5, 20°C
0.0062
S-(2-nitrophenyl)-L-cysteine
-
pH 8.0, 25°C, recombinant mutant F449A
3.3
S-(2-nitrophenyl)-L-cysteine
-
pH 8.0, 25°C, recombinant mutant F448H
3.9
S-(2-nitrophenyl)-L-cysteine
-
recombinant mutant S51A, pH 8.0, 30°C
4.5
S-(2-nitrophenyl)-L-cysteine
-
pH 8.0, 25°C, recombinant mutant F448L
9.6
S-(2-nitrophenyl)-L-cysteine
-
pH 8.0, 25°C, recombinant mutant F448A
46
S-(2-nitrophenyl)-L-cysteine
-
pH 8.0, 25°C, recombinant wild-type enzyme
46
S-(2-nitrophenyl)-L-cysteine
-
recombinant wild-type enzyme, pH 8.0, 30°C
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3
L-Asp
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
3.5
L-Asp
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using L-Tyr as a substrate; wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using S-(o-nitrophenyl)-L-Cys as a substrate
3.6
L-Asp
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
5
L-Glu
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using L-Tyr as a substrate; wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using S-(o-nitrophenyl)-L-Cys as a substrate
66
L-Glu
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
100
L-Glu
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
1.33
L-homoserine
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using L-Tyr as a substrate; wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using S-(o-nitrophenyl)-L-Cys as a substrate
69
L-homoserine
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
82
L-homoserine
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
0.73
L-Met
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using L-Tyr as a substrate; wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using S-(o-nitrophenyl)-L-Cys as a substrate
4.2
L-Met
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
11
L-Met
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
1.7
L-Phe
-
wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using L-Tyr as a substrate; wild type enzyme, in 50 mM potassium phosphate pH 8.0, at 25°C, using S-(o-nitrophenyl)-L-Cys as a substrate
17
L-Phe
-
mutant enzyme D214N, in 50 mM potassium phosphate pH 8.0, at 25°C
22
L-Phe
-
mutant enzyme D214A, in 50 mM potassium phosphate pH 8.0, at 25°C
additional information
additional information
-
rapid-scanning stopped-flow experiments, steady-state and pre-steady-state inhibition kinetics
-
additional information
additional information
-
rapid scanning stopped-flow kinetic experiments, kinetics
-
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0022
-
in strain S12TPL3-delta-hpd mutant (4-hydroxyphenylpyruvate dioxygenase knockout), harvested in logarithmic phase
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8.2
8.5
8
-
recombinant enzyme, L-DOPA synthesis in recombinant Escherichia coli cells
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.5 - 9.2
Symbiobacterium sp.
-
pH 6.5: about 45% of maximal activity, pH 9.2: about 40% of maximal activity
7 - 9.5
Aeromonas phenologenes
-
pH 7.0: about 25% of maximal activity, pH 9.5: about 60% of maximal activity
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
15
-
recombinant enzyme, L-DOPA synthesis in recombinant Escherichia coli cells
30
45
80
45
-
mutant enzyme T15A, using 3,4-dihydroxyphenyl-L-alanine as a substrate
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37 - 75
Symbiobacterium sp.
-
37°C: about 65% of maximal activity, 75°C: about 45% of maximal activity
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pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131
-
UniProt
brenda
Symbiobacterium sp.
-
33441, 33442, 33455, 33469, 650039, 650404, 651062, 652265, 653702, 653815, 664227, 664834, 665928, 678212, 678401, 679758, 713693, 713730, 726642, 726700, 727663
-
-
brenda
-
UniProt
brenda
wild type enzyme and mutant enzymes R381A, R381V and R381I
UniProt
brenda
Escherichia intermedia A-21; formerly Escherichia intermedia
-
-
brenda
SV370; wild-type enzyme, mutant enzyme R100T, mutant enzyme R100T/V283R
-
-
brenda
-
-
-
brenda
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
wild-type S12 and its mutant strains S12TPL3 (optimized for phenol production)
brenda
additional information
additional information
-
optimization and evaluation of cultivation conditions, overview
brenda
additional information
-
optimization and evaluation of cultivation conditions, overview
-
brenda
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
-
the carbon-carbon lyases, tryptophan indole lyase (TIL, EC 4.1.99.1) and tyrosine phenol-lyase (TPL) are bacterial enzymes which catalyze the reversible elimination of indole and phenol from L-tryptophan and L-tyrosine, respectively. These pyridoxal 5'-phosphate-dependent enzymes show high sequence homology (about 40% identity) and both form homotetrameric structures. Pre-steady state kinetics with TPL and TIL show rapid formation of external aldimine intermediates, followed by deprotonation to give quinonoid intermediates absorbing at about 500 nm. The active sites of TIL and TPL are highly conserved with the exceptions of these residues: Arg381(TPL)/Ile396 (TIL), Thr124 (TPL)/Asp137 (TIL), and Phe448 (TPL)/His463 (TIL). The conserved tyrosine, Tyr71 (TPL)/Tyr74 (TIL) is essential for elimination activity with both enzymes, and likely plays a role as a proton donor to the leaving group.A unique feature of TIL and TPL is another strictly conserved lysine (Lys256 in Citrobacter freundii TPL) immediately preceding the pyridoxal 5'-phosphate-binding lysine, and hydrogen bonded to a water molecule bound to the monovalent cation. The mechanisms of TPL and TIL require both substrate strain and acid/base catalysis, and substrate strain is probably responsible for the very high substrate specificity of TPL and TIL. Both enzymes require a monovalent cation, either K+, NH4+, Rb+ or Cs+ for activity, with Na+ and Li+ giving little or no activity. The active site residues involved in PLP binding are highly conserved for both TIL and TPL. Sequence comparisons
metabolism
-
tyrosine phenol lyase catalyses the biosynthesis of L-tyrosine/L-DOPA from ammonium, pyruvate and phenolic substrates through the reverse of alpha- and beta-elimination reaction
physiological function
-
hyper-3,4-dihydroxyphenyl-L-alanine-producing strain by a mutant TyrR, an activator of tpl, whereby highest productivity is obtained for the strain grown under non-induced conditions, which is 30fold higher than that obtained for tyrosine-induced wild-type cells
additional information
additional information
-
the Ser51 residue interacts with the side chain amino group of Lys257 at the stage of C-alpha-proton abstraction. This interaction ensures the correct orientation of the side chain of Lys257 accepting the C-alpha-proton of the external aldimine and stabilizes its ammonium form. Ser51 takes part in formation of a chain of hydrogen bonds which is necessary to perform the transfer of the Calpha-proton to the C-4'-position of the leaving phenol group in the reaction with the natural substrate. Ser51 is necessary for efficient C-alpha-proton abstraction and its following transfer to the 4'-position of the leaving phenol group in the alpha,beta-elimination reaction of L-tyrosine
additional information
-
essential catalytic residues are Tyr71, Thr124, Arg381, and Phe448. There are two quinonoid structures of the enzyme, relaxed and tense. In the relaxed structure, the substrate aromatic ring is in plane with the Cbeta-Cgamma bond, but in the tense structure, the substrate aromatic ring is about 20° out of plane with the Cbeta-Cgamma bond. In the tense structure, hydrogen bonds are formed between the substrate OH and the guanidinium of Arg381 and the OH of Thr124, and the phenyl rings of Phe448 and 449 provide steric strain. Structure of the active site of the TPL quinonoid complex with L-alanine and pyridine N-oxide, overview
Show AA Sequence and Transmembrane Helices (602 entries)
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
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PDB
SCOP
CATH
UNIPROT
ORGANISM
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
52000
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homotetramer
tetramer
homotetramer
-
TPL is a homotetramer with four active sites, two per catalytic dimer, located at the monomer-monomer interface
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CRYSTALLIZATION/commentary
ORGANISM
UNIPROT
LITERATURE
analysis of crystal structure with bound competitive inhibitor 4-hydroxyphenylpropionate, PDB ID 1AX4
-
hanging drop vapour diffusion method with 50 mM potassium phosphate (pH 8.0), 2 mM DTT, 0.2 M KCl, and 32.5% (w/v) PEG2000 for the apo-enzyme, and 50 mM triethanolamine buffer (pH 8.0), containing 0.5 mM PLP, 2 mM DTT, 0.4-0.8 M KCl, and 35-38% (w/v) PEG5000 for the holo-enzyme
-
in complex with L-methionine and L-phenylalanine, wild type and mutant Y71F
-
purified wild-type enzyme in ternary complex with pyridine N-oxide and the L-Ala quinonoid reaction intermediate by soaking wild-type TPL crystals in the stabilization solution containing 40% w/v PEG 5000 MME, 50 mM triethanolamine, pH 8.0, 0.25 M KCl, 0.2 mM pyridoxal 5'-phosphate, 0.5 mM dithiothreitol, with 100 mM L-Ala and a saturating concentration of pyridine N-oxide for 20 s, and purified Y71F or F448H mutant enzymes with bound substrate in quinoid intermediate state by soaking the crystals in the same stabilizing solution but with addition of 10 mM 3-fluoro-L-Tyr for 30 s, X-ray diffraction structure determination and analysis at 2.05-2.25 A resolution
-
the quinonoid intermediates of tyrosine phenol-lyase with L-alanine and L-methionine are trapped in the crystalline state and their structures are determined at 1.9 A and 1.95 A resolution, respectively
-
tyrosine phenol-lyase complexed with 3-(4'-hydroxyphenyl)propionic acid
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D214A
-
mutant does not catalyze the decomposition of L-Tyr and 3-fluoro-L-Tyr
D214N
-
mutant does not catalyze the decomposition of L-Tyr and 3-fluoro-L-Tyr
E69D/K256R
-
no activity with L-Tyr, turnover-number for S-(o-nitrophenyl)-L-cysteine is 1.5fold lower than the wild-type value, turnover-number for S-ethyl-L-Cys is 11fold lower than the wild-type value
F448A
-
site-directed mutagenesis, the mutant shows a 104fold reduced activity with L-tyrosine compared to wild-type, stopped-flow kinetics of enzyme mutant F448A. The mutant F448A TPL forms quinonoid intermediates from L-tyrosine and S-ethyl-L-cysteine with rate constants similar to those of wild-type TPL, and can form an aminoacrylate intermediate from S-ethyl-L-cysteine but not L-tyrosine, with a rate constant similar to that of wild-type TPL
F448H
F448L
-
site-directed mutagenesis, the mutant shows a 103fold reduced activity with L-tyrosine compared to wild-type
F449A
-
site-directed mutagenesis, the mutant shows a 104fold reduced activity with L-tyrosine compared to wild-type
H343A
K256A
-
turnover-number for L-Tyr is 3500fold lower than wild-type value, turnover-number for S-(o-nitrophenyl)-L-cysteine is 560fold lower than the wild-type value, turnover-number for S-ethyl-L-Cys is 1560fold lower than the wild-type value, activity is not increased by addition of monovalent cations, K+, Na+, Li+, Rb+, or NH4+
K256H
-
turnover-number for L-Tyr is 26923fold lower than wild-type value, turnover-number for S-(o-nitrophenyl)-L-cysteine is 189fold lower than the wild-type value, turnover-number for S-ethyl-L-Cys is 443fold lower than the wild-type value, activity is not increased by addition of monovalent cations, K+, Na+, Li+, Rb+, or NH4+
K256R
-
turnover-number for L-Tyr is 29fold lower than wild-type value, turnover-number for S-(o-nitrophenyl)-L-cysteine is 30fold lower than the wild-type value, turnover-number for S-ethyl-L-Cys is 195fold lower than the wild-type value
M379V
-
changed substrate specificity; site-directed mutagenesis, the mutant is active with o-cresol, o-methoxyphenol, and o-chlorophenol, as well as 3-methyl-, 3-methoxy-, 3 -fluoro, and 3-chloro-L-tyrosine in contrast to the wild-type enzyme
N185A
-
2% residual activtiy with L-tyrosine or 3-fluoro-L-tyrosine, N185 stabilizes reaction intermediate
R381A
R381I
R381V
S51A
-
site-directed mutagenesis, the mutation leads to a decrease of the kcat/Km parameter for reactions with L-tyrosine and 3-fluoro-L-tyrosine by three orders of magnitude, compared to the wild-type enzyme, phenotype, overview. Influence of replacement of Ser51 by Ala on the kinetic parameters of TPL reactions with inhibitory L-phenylalanine and L-methionine, kinetics and structures, overview
T124A
T124D
T124D/F448H
-
very little activity with L-tyrosine, significant activity with S-(o-nitrophenyl)-L-cysteine, S-alkyl-L-cysteine and beta-chloro-L-alanine
T15A
-
exhibits a 2fold improved activity towards 3,4-dihydroxyphenyl-L-alanine
Y71F
R100T/V283R
A13V
-
mutant exhibits higher temperature and denaturant stability than wild-type enzyme
T129I/A13V
-
increases the activity and thermal stability of the enzyme
T129I/A13V/E83K/T407A
-
increases the activity and thermal stability of the enzyme
T451A/A13V/E83K
-
increases the activity and thermal stability of the enzyme
T451A/A13V/E83K/T407A
-
increases the activity and thermal stability of the enzyme
additional information
F448H
-
very little activity with L-tyrosine, significant activity with S-(o-nitrophenyl)-L-cysteine, S-alkyl-L-cysteine and beta-chloro-L-alanine
F448H
-
very low activity with L-tyrosine, reduced activity with other substrates
F448H
-
site-directed mutagenesis, the mutant enzyme lacks the beta-elimination activity with L-Tyr or 3-fluoro-L-tyrosine as substrates, instead, it accumulates quinonoid intermediate
F448H
-
site-directed mutagenesis, enzyme mutant crystal structure with bound 3-fluoro-L-tyrosine, tense and closed conformation of F448H TPL quinonoid complex with the ligand, overview. Mutant F448H TPL has very low catalytic activity with L-tyrosine compared to wild-type
F448H
-
site-directed mutagenesis, enzyme mutant crystal structure with bound 3-fluoro-L-tyrosine, tense and closed conformation of F448H TPL quinonoid complex with the ligand, overview
H343A
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all substrates for alpha,beta-elimination, except S-ethyl-L-Cys, exhibit lower turnover number values with the mutant enzyme than with the wild-type enzyme. The mutant shows slower rates of deuterium isotope exchange for L-Phe and L-Met than does the wild type enzyme. The turnover-number for 3-fluoro-L-Tyr is pH-dependent for the mutant enzyme, whereas it is pH-independent for the wild-type enzyme. His343 does play an important function in catalysis, possibly by facilitating the conformational change from an ‘open' to ‘closed' form when substrates bind
H343A
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site-directed mutagenesis, the mutant shows altered kinetics compared to wild-type enzyme
R381A
beta-elimination activity has been reduced by 0.0001fold compared to wild type enzyme
R381A
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dramatic decrease in activity with L-tyrosine, but little effect on activity with other substrates
R381A
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site-directed mutagenesis, the mutant shows very low remaining activity compared to wild-type
R381I
no detectable beta-elimination activity with L-Tyr as substrate
R381I
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dramatic decrease in activity with L-tyrosine, but little effect on activity with other substrates
R381V
no detectable beta-elimination activity with L-Tyr as substrate
R381V
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dramatic decrease in activity with L-tyrosine, but little effect on activity with other substrates
T124A
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very little activity with L-tyrosine, significant activity with S-(o-nitrophenyl)-L-cysteine, S-alkyl-L-cysteine and beta-chloro-L-alanine
T124A
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site-directed mutagenesis, the mutant shows very low remaining activity compared to wild-type