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Literature summary for 1.14.17.3 extracted from

  • Bell, J.; El Meskini, R.; D'Amato, D.; Mains, R.E.; Eipper, B.A.
    Mechanistic investigation of peptidylglycine alpha-hydroxylating monooxygenase via intrinsic tryptophan fluorescence and mutagenesis (2003), Biochemistry, 42, 7133-7142.
    View publication on PubMed
Search for more literature for 1.14.17.3

Cloned(Commentary)

Cloned (Comment) Organism
transient expression of wild-type and mutant enzymes in HEK293 cells, stable expression of wild-type and mutant enzymes in CHO cells Rattus norvegicus

Protein Variants

Protein Variants Comment Organism
Q170A site-directed, PCR-based mutagenesis, altered Km compared to the wild-type enyme Rattus norvegicus
Q170E site-directed, PCR-based mutagenesis, kinetics similar to the wild-type enyme Rattus norvegicus
Q170L site-directed, PCR-based mutagenesis, kinetics similar to the wild-type enyme Rattus norvegicus
Q170N site-directed, PCR-based mutagenesis, kinetics similar to the wild-type enyme Rattus norvegicus
Y79W site-directed, PCR-based mutagenesis, altered kinetics compared to the wild-type enzyme, highly reduced activity Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information fluorescence spectroscopic analysis of intrinsic tryptophan Rattus norvegicus
0.0012
-
 N-acetyl-Tyr-Val-Gly purified recombinant wild-type enzyme, pH 7.0 Rattus norvegicus
0.0023
-
 N-acetyl-Tyr-Val-Gly purified recombinant mutant Y79W, pH 7.0 Rattus norvegicus
0.0029
-
 N-acetyl-Tyr-Val-Gly purified recombinant mutant Q170A, pH 7.0 Rattus norvegicus
0.0063
-
 N-acetyl-Tyr-Val-Gly purified recombinant wild-type enzyme, pH 5.5 Rattus norvegicus
0.0065
-
 N-acetyl-Tyr-Val-Gly purified recombinant mutant Q170A, pH 5.5 Rattus norvegicus
0.009
-
 peptidylglycine transiently expressed mutants Q170N and Q170L, pH 5.5 Rattus norvegicus
0.0098
-
 N-acetyl-Tyr-Val-Gly purified recombinant mutant Y79W, pH 5.5 Rattus norvegicus
0.01
-
 peptidylglycine transiently expressed mutant Q170E, pH 5.5 Rattus norvegicus
0.011
-
 peptidylglycine transiently expressed wild-type enzyme, pH 5.5 Rattus norvegicus
0.018
-
 peptidylglycine transiently expressed mutants Q170A and Y79W, pH 5.5 Rattus norvegicus

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ dependent on, metalloenzyme, binding analysis Rattus norvegicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
peptidylglycine + ascorbate + O2 Rattus norvegicus
-
 peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
-
 ?

Organism

Organism UniProt Comment Textmining
Rattus norvegicus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from CHO cells by a 3-step chromatographic method Rattus norvegicus

Reaction

Reaction Comment Organism Reaction ID
[peptide]-glycine + 2 ascorbate + O2 = [peptide]-(2S)-2-hydroxyglycine + 2 monodehydroascorbate + H2O mechanism, substrate binding structure Rattus norvegicus
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
N-acetyl-Tyr-Val-Gly + ascorbate + O2
-
 Rattus norvegicus N-acetyl-Tyr-Val-(2-hydroxyglycine) + dehydroascorbate + H2O
-
 ?
peptidylglycine + ascorbate + O2
-
 Rattus norvegicus peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
-
 ?
peptidylglycine + ascorbate + O2 2 Cu2+, A and B, are involved in the catalytic reaction Rattus norvegicus peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
-
 ?
Phe-Gly-Phe-Gly + ascorbate + O2
-
 Rattus norvegicus Phe-Gly-Phe-(2-hydroxyglycine) + dehydroascorbate + H2O
-
 ?

Synonyms

Synonyms Comment Organism
peptidylglycine alpha-hydroxylating monooxygenase
-
 Rattus norvegicus
PHM
-
 Rattus norvegicus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0035
-
 N-acetyl-Tyr-Val-Gly purified recombinant mutant Y79W, pH 7.0 Rattus norvegicus
0.09
-
 N-acetyl-Tyr-Val-Gly purified recombinant mutant Y79W, pH 5.5 Rattus norvegicus
0.53
-
 N-acetyl-Tyr-Val-Gly purified recombinant wild-type enzyme, pH 7.0 Rattus norvegicus
0.62
-
 N-acetyl-Tyr-Val-Gly purified recombinant mutant Q170A, pH 7.0 Rattus norvegicus
14
-
 N-acetyl-Tyr-Val-Gly purified recombinant mutant Q170A, pH 5.5 Rattus norvegicus
15
-
 N-acetyl-Tyr-Val-Gly purified recombinant wild-type enzyme, pH 5.5 Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
-
 Rattus norvegicus

Cofactor

Cofactor Comment Organism Structure
ascorbate dependent on Rattus norvegicus