1.13.11.27 additional information - additional information - 395368, 395376 1.13.11.27 additional information - additional information Kd value for 4-hydroxyphenylpyruvate with 4-hydroxyphenylpyruvate dioxygenase Fe2+ is 29 microM, whereas the binding constant for phenylpyruvate is determined to be 319 microM 672170 1.13.11.27 additional information - additional information the Km-value is the same at 25°C and at 38°C 395367, 395377 1.13.11.27 0.0017 - 4-hydroxyphenylpyruvate mutant V218L/A327R/I340E/L359M , pH 7, 25°C 765506 1.13.11.27 0.00187 - 4-hydroxyphenylpyruvate wild-type, pH 7, 25°C 765756 1.13.11.27 0.002 - 4-hydroxyphenylpyruvate mutant V218I/A327R/I340E/L359M, pH 7, 25°C 765506 1.13.11.27 0.0026 - 4-hydroxyphenylpyruvate mutant A327R/L359M, pH 7, 25°C 765506 1.13.11.27 0.00304 - 4-hydroxyphenylpyruvate pH 7, 25°C 765506 1.13.11.27 0.0031 - 4-hydroxyphenylpyruvate mutant V218I/A327R/L359M, pH 7, 25°C 765506 1.13.11.27 0.0043 - 4-hydroxyphenylpyruvate - 395379 1.13.11.27 0.0043 - 4-hydroxyphenylpyruvate mutant L359M, pH 7, 25°C 765506 1.13.11.27 0.0048 - 4-hydroxyphenylpyruvate wild-type, pH 7, 25°C 765506 1.13.11.27 0.0053 - 4-hydroxyphenylpyruvate mutant S267A, pH 7, 25°C 765756 1.13.11.27 0.0069 - 4-hydroxyphenylpyruvate activity is measured polarographically, assays include 0.5 microM of enzyme, 500 microM betaME, Fe2+ stoichiometric with enzyme, and 400 microM 4-hydroxyophenylpyruvate in 20 mM HEPES at pH 7.0 at 4°C 672170 1.13.11.27 0.0075 - 4-hydroxyphenylpyruvate recombinant wild-type enzyme, pH and temperature not specified in the publication 725432 1.13.11.27 0.0087 - 4-hydroxyphenylpyruvate pH 7.0, 25°C, mutant P243T 724325 1.13.11.27 0.009 - 4-hydroxyphenylpyruvate recombinant mutant S246A, pH and temperature not specified in the publication 725432 1.13.11.27 0.0091 - 4-hydroxyphenylpyruvate mutant F381A, pH 7, 25°C 765756 1.13.11.27 0.0108 - 4-hydroxyphenylpyruvate mutant N282A, pH 7, 25°C 765756 1.13.11.27 0.0143 - 4-hydroxyphenylpyruvate mutant V218I/A327R/I340E/L359M/G411A, pH 7, 25°C 765506 1.13.11.27 0.015 - 4-hydroxyphenylpyruvate pH 7.0, 25°C, mutant N245Q 724325 1.13.11.27 0.0157 - 4-hydroxyphenylpyruvate mutant N423A, pH 7, 25°C 765756 1.13.11.27 0.025 - 4-hydroxyphenylpyruvate enzyme form 1 395382 1.13.11.27 0.027 - (4-hydroxyphenyl)-pyruvate pH 7.0, 25°C 657978 1.13.11.27 0.027 - 4-hydroxyphenylpyruvate enzyme form 2 395382 1.13.11.27 0.027 - 4-hydroxyphenylpyruvate pH 7.0, 25°C, wild-type enzyme and mutant N245I 724325 1.13.11.27 0.029 - 4-hydroxyphenylpyruvate enzyme form 3 395382 1.13.11.27 0.03 - 4-hydroxyphenylpyruvate - 395369, 395381 1.13.11.27 0.0303 - 4-hydroxyphenylpyruvate mutant Q307A, pH 7, 25°C 765756 1.13.11.27 0.0309 - 4-hydroxyphenylpyruvate mutant G413W, pH 7, 25°C 765506 1.13.11.27 0.0389 - 4-hydroxyphenylpyruvate mutant G411A, pH 7, 25°C 765506 1.13.11.27 0.04 - 4-hydroxyphenylpyruvate - 395367, 395388 1.13.11.27 0.041 - 4-hydroxyphenylpyruvate - 395392 1.13.11.27 0.0432 - 4-hydroxyphenylpyruvate - 702737 1.13.11.27 0.047 - 4-hydroxyphenylpyruvate pH 7.0, 25°C, mutant S230A 724325 1.13.11.27 0.05 - 3,4-Dihydroxyphenylpyruvate - 395389 1.13.11.27 0.05 - 4-hydroxyphenylpyruvate - 395389 1.13.11.27 0.05 - 4-hydroxytetrafluorophenylpyruvate - 395392 1.13.11.27 0.05 - O2 - 395368, 395381 1.13.11.27 0.06 - phenylpyruvate - 395389 1.13.11.27 0.066 - phenylpyruvate - 395392 1.13.11.27 0.069 - O2 pH 7.0, 25°C 657978 1.13.11.27 0.07 - 4-hydroxyphenylpyruvate pH 6.5, 37°C, mutant DELTAL381 726347 1.13.11.27 0.075 - 4-hydroxyphenylpyruvate - 395396 1.13.11.27 0.077 - 4-hydroxyphenylpyruvate wild-type, pH 7,20°C 765723 1.13.11.27 0.08 - 4-hydroxyphenylpyruvate pH 6.5, 37°C, wild-type enzyme and mutant DELTAG388 726347 1.13.11.27 0.083 - 4-hydroxyphenylpyruvate pH 6, 35°C 764952 1.13.11.27 0.09 - 4-hydroxyphenylpyruvate pH 6.5, 37°C, mutant DELTAL385 726347 1.13.11.27 0.0904 - 4-hydroxyphenylpyruvate mutant Q293A, pH 7, 25°C 765756 1.13.11.27 0.098 - 4-hydroxyphenylpyruvate mutant M335H/P336A/P339T/T341D/E363Q/E426Q, pH 7,20°C 765723 1.13.11.27 0.1 - 4-hydroxyphenylpyruvate mutant K10A, pH 6.5, 37°C 765020 1.13.11.27 0.1 - 4-hydroxyphenylpyruvate mutant lacking 17 N-terminal residues, pH 6.5, 37°C 765020 1.13.11.27 0.1 - O2 - 395389 1.13.11.27 0.11 - 4-hydroxyphenylpyruvate pH 6.5, 37°C, mutant E254D 726347 1.13.11.27 0.115 - oxygen activity is measured polarographically, assays include 0.5 microM of enzyme, 500 microM betaME, Fe2+ stoichiometric with enzyme, and 400 microM 4-hydroxyophenylpyruvate in 20 mM HEPES at pH 7.0 at 4°C 672170 1.13.11.27 0.12 - 4-hydroxyphenylpyruvate pH 6.5, 37°C, mutant DELTAG379 726347 1.13.11.27 0.134 - 4-hydroxyphenylpyruvate mutant M335H/P336A/E363Q, pH 7,20°C 765723 1.13.11.27 0.142 - 4-hydroxyphenylpyruvate pH 7.0, 50°C 727448 1.13.11.27 0.153 - 4-hydroxyphenylpyruvate recombinant mutant N261D, pH and temperature not specified in the publication 725432 1.13.11.27 0.18 - 4-hydroxyphenylpyruvate mutant E12A, pH 6.5, 37°C 765020 1.13.11.27 0.18 - 4-hydroxyphenylpyruvate mutant P11A, pH 6.5, 37°C 765020 1.13.11.27 0.2 - 4-hydroxyphenylpyruvate mutant Q334N, pH 6.5, 37°C 764109 1.13.11.27 0.2 - 4-hydroxyphenylpyruvate wild-type, pH 6.5, 37°C 764109 1.13.11.27 0.24 - 4-hydroxyphenylpyruvate pH 6.5, 37°C, mutant R378K 726347 1.13.11.27 0.24 - 4-hydroxyphenylpyruvate wild-type, pH 6.5, 37°C 765020 1.13.11.27 0.25 - 3-thienylpyruvate - 395392 1.13.11.27 0.286 - 4-hydroxyphenylpyruvate recombinant mutant Q358E, pH and temperature not specified in the publication 725432 1.13.11.27 0.3 - 4-hydroxyphenylpyruvate mutant N363A, pH 6.5, 37°C 764109 1.13.11.27 0.3 - 4-hydroxyphenylpyruvate mutant Q334A, pH 6.5, 37°C 764109 1.13.11.27 0.3 - 4-hydroxyphenylpyruvate pH 6.5, 37°C, mutant DELTAN380 726347 1.13.11.27 0.32 - 4-hydroxyphenylpyruvate pH 7,20°C 765723 1.13.11.27 0.37 - 4-hydroxyphenylpyruvate mutant lacking 12 N-terminal residues, pH 6.5, 37°C 765020 1.13.11.27 0.4 - 4-hydroxyphenylpyruvate mutant N363Q, pH 6.5, 37°C 764109 1.13.11.27 0.452 - 4-hydroxyphenylpyruvate recombinant mutant Q272E, pH and temperature not specified in the publication 725432 1.13.11.27 0.5 - 2-thienylpyruvate - 395392 1.13.11.27 0.5 - 4-hydroxyphenylpyruvate - 395388 1.13.11.27 0.5 - 4-hydroxyphenylpyruvate mutant N363D, pH 6.5, 37°C 764109 1.13.11.27 0.5 - 4-hydroxyphenylpyruvate mutant Q334A/N363D, pH 6.5, 37°C 764109 1.13.11.27 0.5 - 4-hydroxyphenylpyruvate mutant Q334N/N363Q, pH 6.5, 37°C 764109 1.13.11.27 0.5 - phenylpyruvate - 395369, 395381 1.13.11.27 0.52 - phenylpyruvate - 395384 1.13.11.27 0.551 - 4-hydroxyphenylpyruvate recombinant mutant Q286E, pH and temperature not specified in the publication 725432 1.13.11.27 0.6 - 4-hydroxyphenylpyruvate mutant Q334A/N363Q, pH 6.5, 37°C 764109 1.13.11.27 0.9 - 4-hydroxyphenylpyruvate mutant Q265E, pH 6.5, 37°C 764109 1.13.11.27 11 - 4-hydroxyphenylpyruvate Solvent is H2O. Steady-state analyses of 4-hydroxyphenylpyruvate dioxygenase using proteo-HPP, 2,3,5,6-tetradeutero-HPP, and proteo-HPP in deuterium oxide are carried out in 1 ml assays on a Hasetech oxygen electrode. The assay mixture include ferrous sulfate (10 microM), DTT (1mM), HPPD (500 nM), and HPP (0-1 mM) in 20 mM HEPES, pH (pD) 7.0, under conditions of atmospheric oxygen (350 microM at 5°C) 672033 1.13.11.27 22 - 4-hydroxyphenylpyruvate Solvent is H2O. Steady-state analyses of 4-hydroxyphenylpyruvate dioxygenase using proteo-HPP, 2,3,5,6-tetradeutero-HPP, and proteo-HPP in deuterium oxide are carried out in 1 ml assays on a Hasetech oxygen electrode. The assay mixture include ferrous sulfate (10 microM), DTT (1mM), HPPD (500 nM), and HPP (0-1 mM) in 20 mM HEPES, pH (pD) 7.0, under conditions of atmospheric oxygen (350 microM at 5°C) 672033 1.13.11.27 54 - 4-hydroxyphenylpyruvate Solvent is D2O. Steady-state analyses of 4-hydroxyphenylpyruvate dioxygenase using proteo-HPP, 2,3,5,6-tetradeutero-HPP, and proteo-HPP in deuterium oxide are carried out in 1 ml assays on a Hasetech oxygen electrode. The assay mixture include ferrous sulfate (10 microM), DTT (1mM), HPPD (500 nM), and HPP (0-1 mM) in 20 mM HEPES, pH (pD) 7.0, under conditions of atmospheric oxygen (350 microM at 5°C) 672033