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Literature summary for 1.14.16.1 extracted from

  • Gersting, S.W.; Staudigl, M.; Truger, M.S.; Messing, D.D.; Danecka, M.K.; Sommerhoff, C.P.; Kemter, K.F.; Muntau, A.C.
    Activation of phenylalanine hydroxylase induces positive cooperativity toward the natural cofactor (2010), J. Biol. Chem., 285, 30686-30697.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.14.16.1

Application

Application Comment Organism
molecular biology an automated fluorescence-based continuous real-time PAH activity assay that is faster and more efficient but as precise and accurate as standard methods is developed Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Homo sapiens

Protein Variants

Protein Variants Comment Organism
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) Homo sapiens
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 Homo sapiens
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 Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 L-phenylalanine wild-type, pH 7.3, 25°C, measurement by standard discontinuous PAH activity assay (HPLC and fluorimetric detection): Vmax: 0.495 micromol L-Tyr/min/mg (without L-Phe preincubation), 1.55 micromol L-Tyr/min/mg (with L-Phe preincubation), S0.5: 0.318 mM (without L-Phe preincubation), S0.5: 0.154 mM (with L-Phe preincubation) Homo sapiens
0.008
-
 (6R)-L-erythro-5,6,7,8-tetrahydrobiopterin wild-type, pH 7.3, 25°C, measurement by continuous PAH activity assay (direct in-well fluorescence detection): Vmax: 2.277 micromol L-Tyr/min/mg (without L-Phe preincubation) Homo sapiens
0.024
-
 (6R)-L-erythro-5,6,7,8-tetrahydrobiopterin wild-type, pH 7.3, 25°C, measurement by standard discontinuous PAH activity assay (HPLC and fluorimetric detection): Vmax: 3.425 micromol L-Tyr/min/mg (with L-Phe preincubation) Homo sapiens
0.318
-
 L-phenylalanine wild-type, pH 7.3, 25°C, measurement by continuous PAH activity assay (direct in-well fluorescence detection): Vmax: 2.53 micromol L-Tyr/min/mg (without L-Phe preincubation), 6.59 micromol L-Tyr/min/mg (with L-Phe preincubation) Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-phenylalanine + (6R)-L-erythro-5,6,7,8-tetrahydrobiopterin + O2 an automated fluorescence-based continuous real-time PAH activity assay that is faster and more efficient but as precise and accurate as standard methods is developed. The assay is performed with and without preincubation of the enzyme with 1 mM L-Phe Homo sapiens L-tyrosine + 4a-hydroxy-(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin
-
 ?

Synonyms

Synonyms Comment Organism
PAH
-
 Homo sapiens
phenylalanine hydroxylase
-
 Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
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 Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.3
-
 assay at Homo sapiens

Cofactor

Cofactor Comment Organism Structure
(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin
-
 Homo sapiens