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Literature summary for 1.11.1.5 extracted from

  • Murphy, E.J.; Metcalfe, C.L.; Basran, J.; Moody, P.C.; Raven, E.L.
    Engineering the substrate specificity and reactivity of a heme protein: creation of an ascorbate binding site in cytochrome c peroxidase (2008), Biochemistry, 47, 13933-13941.
    View publication on PubMed
Search for more literature for 1.11.1.5

Cloned(Commentary)

Cloned (Comment) Organism
-
 Saccharomyces cerevisiae

Crystallization (Commentary)

Crystallization (Comment) Organism
structures for mutants N184R, Y36A, W191F, N184R/W191F, Y36A/W191, FY36A/N184R, Y36A/N184R/W191F, Y36A/N184R/W191F-ascorbate complex, no major perturbations compared to the wild type protein Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
N184R the N184R variant introduces potential hydrogen bonding interactions for ascorbate binding Saccharomyces cerevisiae
N184R/W191F site-directed mutagenesis Saccharomyces cerevisiae
W191F site-directed mutagenesis Saccharomyces cerevisiae
Y36A site-directed mutagenesis, Tyr36 directly blocks the equivalent ascorbate binding site in CcP and was therefore replaced with a less bulky residue Saccharomyces cerevisiae
Y36A/N184R site-directed mutagenesis, no significant spectroscopic changes on reaction with stoichiometric or higher amounts of H2O2 are seen Saccharomyces cerevisiae
Y36A/N184R/W191F site-directed mutagenesis, cytochrome c peroxidase enzyme can duplicate the substrate binding properties of ascorbate peroxidase through the introduction of relatively modest structural changes at Tyr36 and Asn184, no evidence for a porphyrin pi-cation radical Saccharomyces cerevisiae
Y36A/W191F site-directed mutagenesis, no significant spectroscopic changes on reaction with stoichiometric or higher amounts of H2O2 are seen Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.31
-
 ascorbate mutant N184R/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
0.45
-
 ascorbate mutant W191, pH 6.0, 25°C Saccharomyces cerevisiae
0.48
-
 ascorbate mutant FY36A/N184R, pH 6.0, 25°C Saccharomyces cerevisiae
0.5
-
 ascorbate mutant Y36A/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
0.71
-
 ascorbate wild-type cytochrome c peroxidase, pH 6.0, 25°C Saccharomyces cerevisiae
1.3
-
 ascorbate mutant Y36A/N184R/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
1.7
-
 ascorbate mutant N184R, pH 6.0, 25°C Saccharomyces cerevisiae
1.7
-
 ascorbate mutant Y36A, pH 6.0, 25°C Saccharomyces cerevisiae
14
-
 guaiacol mutant Y36A/N184R/W191F, pH 6.0, 25°C, guaiacol oxidation is not significantly affected by any of the mutations, including W191F, which is consistent with the idea that aromatic substrates such as guaiacol bind at a separate location close to the delta-heme edge and is clearly indicative of a different electron transfer pathway for the oxidation of these types of aromatic substrate Saccharomyces cerevisiae
16
-
 guaiacol mutant FY36A/N184R, pH 6.0, 25°C Saccharomyces cerevisiae
27
-
 guaiacol mutant N184R, pH 6.0, 25°C Saccharomyces cerevisiae
34
-
 guaiacol mutant N184R/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
36
-
 guaiacol mutant Y36A, pH 6.0, 25°C Saccharomyces cerevisiae
45
-
 guaiacol mutant Y36A/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
53
-
 guaiacol wild-type cytochrome c peroxidase, pH 6.0, 25°C Saccharomyces cerevisiae
57
-
 guaiacol mutant W191, pH 6.0, 25°C Saccharomyces cerevisiae
93
-
 cytochrome c wild-type cytochrome c peroxidase, pH 6.0, 25°C Saccharomyces cerevisiae
100
-
 cytochrome c mutant W191F, pH 6.0, 25°C, the W191F mutation dramatically reduces the activity toward cytochrome c, but in the other variants which do not contain the W191F mutation the activity toward cytochrome c is largely unaffected Saccharomyces cerevisiae
110
-
 cytochrome c mutant FY36A/N184R, pH 6.0, 25°C Saccharomyces cerevisiae
140
-
 cytochrome c mutant N184R, pH 6.0, 25°C Saccharomyces cerevisiae
160
-
 cytochrome c mutant Y36A/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
230
-
 cytochrome c mutant Y36A, pH 6.0, 25°C Saccharomyces cerevisiae
300
-
 cytochrome c mutant Y36A/N184R/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
670
-
 cytochrome c mutant N184R/W191F, pH 6.0, 25°C Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ferrocytochrome c + H2O2 Saccharomyces cerevisiae
-
 ferricytochrome c + H2O
-
 ?

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae P00431 baker’s yeast
-

Purification (Commentary)

Purification (Comment) Organism
-
 Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ascorbate + H2O2
-
 Saccharomyces cerevisiae dehydroascorbate + H2O
-
 ?
cytochrome c + H2O2
-
 Saccharomyces cerevisiae ?
-
 ?
ferrocytochrome c + H2O2
-
 Saccharomyces cerevisiae ferricytochrome c + H2O
-
 ?
guaiacol + H2O2
-
 Saccharomyces cerevisiae 2-methoxy-cyclohexa-2,5-dienone + H2O
-
 ?

Synonyms

Synonyms Comment Organism
cytochrome c peroxidase
-
 Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.06
-
 cytochrome c mutant Y36A/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
0.08
-
 cytochrome c mutant N184R/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
0.25
-
 ascorbate mutant W191, pH 6.0, 25°C Saccharomyces cerevisiae
0.27
-
 ascorbate mutant N184R/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
0.45
-
 ascorbate mutant Y36A/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
0.66
-
 ascorbate mutant FY36A/N184R, pH 6.0, 25°C Saccharomyces cerevisiae
0.83
-
 ascorbate wild-type cytochrome c peroxidase, pH 6.0, 25°C Saccharomyces cerevisiae
0.9
-
 guaiacol mutant FY36A/N184R, pH 6.0, 25°C Saccharomyces cerevisiae
1.3
-
 ascorbate mutant Y36A, pH 6.0, 25°C Saccharomyces cerevisiae
1.5
-
 ascorbate mutant N184R, pH 6.0, 25°C Saccharomyces cerevisiae
1.5
-
 guaiacol mutant Y36A/N184R/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
1.6
-
 cytochrome c mutant Y36A/N184R/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
1.7
-
 cytochrome c mutant W191, pH 6.0, 25°C Saccharomyces cerevisiae
2.6
-
 ascorbate mutant Y36A/N184R/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
3
-
 guaiacol mutant N184R, pH 6.0, 25°C Saccharomyces cerevisiae
3.2
-
 guaiacol mutant Y36A/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
4.1
-
 guaiacol wild-type cytochrome c peroxidase, pH 6.0, 25°C Saccharomyces cerevisiae
4.9
-
 guaiacol mutant N184R/W191F, pH 6.0, 25°C Saccharomyces cerevisiae
5.4
-
 guaiacol mutant Y36A, pH 6.0, 25°C Saccharomyces cerevisiae
14
-
 guaiacol mutant W191, pH 6.0, 25°C Saccharomyces cerevisiae
570
-
 cytochrome c mutant N184R, pH 6.0, 25°C Saccharomyces cerevisiae
580
-
 cytochrome c mutant Y36A, pH 6.0, 25°C Saccharomyces cerevisiae
600
-
 cytochrome c mutant FY36A/N184R, pH 6.0, 25°C Saccharomyces cerevisiae
1510
-
 cytochrome c wild-type cytochrome c peroxidase, pH 6.0, 25°C Saccharomyces cerevisiae