BRENDA - Enzyme Database

Role of arginine 177 in the MnII binding site of manganese peroxidase. Studies with R177D, R177E, R177N, and R177Q mutants

Gelpke, M.D.S.; Youngs, H.L.; Gold, M.H.; Eur. J. Biochem. 267, 7038-7045 (2000)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.11.1.13
malonate
-
Phanerochaete chrysosporium
1.11.1.13
oxalate
activates by chelating and stabilizing Mn3+
Phanerochaete chrysosporium
Cloned(Commentary)
EC Number
Commentary
Organism
1.11.1.13
cDNA sequences of several MnP-encoding genes, including mnp1, mnp2 and mnp3
Phanerochaete chrysosporium
Crystallization (Commentary)
EC Number
Crystallization
Organism
1.11.1.13
-
Phanerochaete chrysosporium
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.11.1.13
E35Q
engineered mutant
Phanerochaete chrysosporium
1.11.1.13
R177A
mutant with reduced binding efficiency for Mn2+: disruption in the salt-bridge between Arg-177 and the Mn2+ binding ligand Glu-35
Phanerochaete chrysosporium
1.11.1.13
R177D
mutant with decreased electron-transfer rate and reduced binding efficiency for Mn2+: disruption in the salt-bridge between Arg-177 and the Mn2+ binding ligand Glu-35, higher redox potential for the enzyme-bound Mn2+
Phanerochaete chrysosporium
1.11.1.13
R177E
mutant with decreased electron-transfer rate and reduced binding efficiency for Mn2+: disruption in the salt-bridge between Arg-177 and the Mn2+ binding ligand Glu-35, higher redox potential for the enzyme-bound Mn2+
Phanerochaete chrysosporium
1.11.1.13
R177K
mutant with reduced binding efficiency for Mn2+: disruption in the salt-bridge between Arg-177 and the Mn2+ binding ligand Glu-35
Phanerochaete chrysosporium
1.11.1.13
R177N
mutant with decreased electron-transfer rate and reduced binding efficiency for Mn2+: disruption in the salt-bridge between Arg-177 and the Mn2+ binding ligand Glu-35, higher redox potential for the enzyme-bound Mn2+
Phanerochaete chrysosporium
1.11.1.13
R177Q
mutant with decreased electron-transfer rate and reduced binding efficiency for Mn2+: disruption in the salt-bridge between Arg-177 and the Mn2+ binding ligand Glu-35, higher redox potential for the enzyme-bound Mn2+
Phanerochaete chrysosporium
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.11.1.13
0.0057
-
H2O2
mutant R177E
Phanerochaete chrysosporium
1.11.1.13
0.0078
-
H2O2
mutant R177Q
Phanerochaete chrysosporium
1.11.1.13
0.0121
-
H2O2
mutant R177N
Phanerochaete chrysosporium
1.11.1.13
0.0126
-
H2O2
mutant R177D
Phanerochaete chrysosporium
1.11.1.13
0.039
-
H2O2
mutant R177A
Phanerochaete chrysosporium
1.11.1.13
0.044
-
H2O2
mutant R177K
Phanerochaete chrysosporium
1.11.1.13
0.055
-
H2O2
wild-type MnP
Phanerochaete chrysosporium
1.11.1.13
0.09
-
Mn2+
wild-type MnP
Phanerochaete chrysosporium
1.11.1.13
1.64
-
Mn2+
mutant R177A
Phanerochaete chrysosporium
1.11.1.13
2.15
-
Mn2+
mutant R177D
Phanerochaete chrysosporium
1.11.1.13
2.32
-
Mn2+
mutant R177K
Phanerochaete chrysosporium
1.11.1.13
2.91
-
Mn2+
mutant R177Q
Phanerochaete chrysosporium
1.11.1.13
3.61
-
Mn2+
mutant R177N
Phanerochaete chrysosporium
1.11.1.13
3.9
-
Mn2+
mutant R177E
Phanerochaete chrysosporium
1.11.1.13
4.4
-
Mn2+
mutant E35Q
Phanerochaete chrysosporium
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.11.1.13
extracellular
-
Phanerochaete chrysosporium
-
-
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.11.1.13
46000
-
x * 46000, wild-type and mutants R177D, R177E, R177N and R177Q, SDS-PAGE
Phanerochaete chrysosporium
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.11.1.13
Mn2+ + H+ + H2O2
Phanerochaete chrysosporium
important component of lignin degradation system
Mn3+ + H2O
-
Phanerochaete chrysosporium
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.11.1.13
Phanerochaete chrysosporium
-
; auxotrophic strain OGC107-1; MnP1; white rot basidomycete
-
1.11.1.13
Phanerochaete chrysosporium OGC101
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.11.1.13
purification of the mutant enzymes R177D, R177E, R177N and R177Q
Phanerochaete chrysosporium
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.11.1.13
Mn2+ + H+ + H2O2
unique binding and oxidation site for Mn2+, single Mn atom is hexacoordinate, with two water ligands and four carboxylate ligands from heme propionate 6 and amino acids Glu-35, Glu-39 and Asp-179
439839
Phanerochaete chrysosporium
Mn3+ + H2O
freely diffusible, enzyme-generated Mn(III)-organic-acid complex oxidizes phenolic substrates
439839
Phanerochaete chrysosporium
-
1.11.1.13
Mn2+ + H+ + H2O2
unique binding and oxidation site for Mn2+, single Mn atom is hexacoordinate, with two water ligands and four carboxylate ligands from heme propionate 6 and amino acids Glu-35, Glu-39 and Asp-179
439839
Phanerochaete chrysosporium
Mn3+ + H2O
Mn3+ oxidizes lignin
439839
Phanerochaete chrysosporium
-
1.11.1.13
Mn2+ + H+ + H2O2
role for Arg-177 in promoting efficient Mn2+ binding and oxidation by MnP
439839
Phanerochaete chrysosporium
Mn3+ + H2O
freely diffusible, enzyme-generated Mn(III)-organic-acid complex oxidizes phenolic substrates
439839
Phanerochaete chrysosporium
-
1.11.1.13
Mn2+ + H+ + H2O2
role for Arg-177 in promoting efficient Mn2+ binding and oxidation by MnP
439839
Phanerochaete chrysosporium
Mn3+ + H2O
Mn3+ oxidizes lignin
439839
Phanerochaete chrysosporium
-
1.11.1.13
Mn2+ + H+ + H2O2
important component of lignin degradation system
439839
Phanerochaete chrysosporium
Mn3+ + H2O
-
439839
Phanerochaete chrysosporium
?
1.11.1.13
Mn2+ + H+ + H2O2
unique binding and oxidation site for Mn2+, single Mn atom is hexacoordinate, with two water ligands and four carboxylate ligands from heme propionate 6 and amino acids Glu-35, Glu-39 and Asp-179
439839
Phanerochaete chrysosporium OGC101
Mn3+ + H2O
freely diffusible, enzyme-generated Mn(III)-organic-acid complex oxidizes phenolic substrates
439839
Phanerochaete chrysosporium OGC101
-
1.11.1.13
additional information
structural properties
439839
Phanerochaete chrysosporium
?
-
-
-
-
1.11.1.13
additional information
catalytic cycle with oxidized intermediates MnP compound I and II
439839
Phanerochaete chrysosporium
?
-
-
-
-
1.11.1.13
additional information
enzyme oxidizes ferrocyanide
439839
Phanerochaete chrysosporium
?
-
-
-
-
1.11.1.13
additional information
enzyme oxidizes bromide
439839
Phanerochaete chrysosporium
?
-
-
-
-
1.11.1.13
additional information
structural properties
439839
Phanerochaete chrysosporium OGC101
?
-
-
-
-
1.11.1.13
additional information
catalytic cycle with oxidized intermediates MnP compound I and II
439839
Phanerochaete chrysosporium OGC101
?
-
-
-
-
1.11.1.13
additional information
enzyme oxidizes ferrocyanide
439839
Phanerochaete chrysosporium OGC101
?
-
-
-
-
1.11.1.13
additional information
enzyme oxidizes bromide
439839
Phanerochaete chrysosporium OGC101
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
1.11.1.13
?
x * 46000, wild-type and mutants R177D, R177E, R177N and R177Q, SDS-PAGE
Phanerochaete chrysosporium
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.11.1.13
0.77
-
Mn2+
mutant E35Q
Phanerochaete chrysosporium
1.11.1.13
17.3
-
H2O2
mutant R177E
Phanerochaete chrysosporium
1.11.1.13
23.2
-
H2O2
mutant R177Q
Phanerochaete chrysosporium
1.11.1.13
25
-
Mn2+
mutant R177Q
Phanerochaete chrysosporium
1.11.1.13
25.5
-
Mn2+
mutant R177E
Phanerochaete chrysosporium
1.11.1.13
28.8
-
H2O2
mutant R177D
Phanerochaete chrysosporium
1.11.1.13
30.2
-
Mn2+
mutant R177D
Phanerochaete chrysosporium
1.11.1.13
34.6
-
Mn2+
mutant R177N
Phanerochaete chrysosporium
1.11.1.13
35.8
-
H2O2
mutant R177N
Phanerochaete chrysosporium
1.11.1.13
184
-
Mn2+
mutant R177A
Phanerochaete chrysosporium
1.11.1.13
190
-
H2O2
mutant R177A
Phanerochaete chrysosporium
1.11.1.13
252
-
H2O2
wild-type MnP
Phanerochaete chrysosporium
1.11.1.13
256
-
Mn2+
wild-type MnP
Phanerochaete chrysosporium
1.11.1.13
262
-
Mn2+
mutant R177K
Phanerochaete chrysosporium
1.11.1.13
273
-
H2O2
mutant R177K
Phanerochaete chrysosporium
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.11.1.13
heme
heme iron of the native enzyme is ferric, high-spin and pentacoordinate with a proximal histidine ligand, important catalytic residues in the heme pocket are conserved: proximal His-173 and Asp-242, distal His-46, Arg-42, Asn-80 and Glu-74
Phanerochaete chrysosporium
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.11.1.13
malonate
-
Phanerochaete chrysosporium
1.11.1.13
oxalate
activates by chelating and stabilizing Mn3+
Phanerochaete chrysosporium
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.11.1.13
cDNA sequences of several MnP-encoding genes, including mnp1, mnp2 and mnp3
Phanerochaete chrysosporium
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.11.1.13
heme
heme iron of the native enzyme is ferric, high-spin and pentacoordinate with a proximal histidine ligand, important catalytic residues in the heme pocket are conserved: proximal His-173 and Asp-242, distal His-46, Arg-42, Asn-80 and Glu-74
Phanerochaete chrysosporium
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.11.1.13
-
Phanerochaete chrysosporium
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.11.1.13
E35Q
engineered mutant
Phanerochaete chrysosporium
1.11.1.13
R177A
mutant with reduced binding efficiency for Mn2+: disruption in the salt-bridge between Arg-177 and the Mn2+ binding ligand Glu-35
Phanerochaete chrysosporium
1.11.1.13
R177D
mutant with decreased electron-transfer rate and reduced binding efficiency for Mn2+: disruption in the salt-bridge between Arg-177 and the Mn2+ binding ligand Glu-35, higher redox potential for the enzyme-bound Mn2+
Phanerochaete chrysosporium
1.11.1.13
R177E
mutant with decreased electron-transfer rate and reduced binding efficiency for Mn2+: disruption in the salt-bridge between Arg-177 and the Mn2+ binding ligand Glu-35, higher redox potential for the enzyme-bound Mn2+
Phanerochaete chrysosporium
1.11.1.13
R177K
mutant with reduced binding efficiency for Mn2+: disruption in the salt-bridge between Arg-177 and the Mn2+ binding ligand Glu-35
Phanerochaete chrysosporium
1.11.1.13
R177N
mutant with decreased electron-transfer rate and reduced binding efficiency for Mn2+: disruption in the salt-bridge between Arg-177 and the Mn2+ binding ligand Glu-35, higher redox potential for the enzyme-bound Mn2+
Phanerochaete chrysosporium
1.11.1.13
R177Q
mutant with decreased electron-transfer rate and reduced binding efficiency for Mn2+: disruption in the salt-bridge between Arg-177 and the Mn2+ binding ligand Glu-35, higher redox potential for the enzyme-bound Mn2+
Phanerochaete chrysosporium
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.11.1.13
0.0057
-
H2O2
mutant R177E
Phanerochaete chrysosporium
1.11.1.13
0.0078
-
H2O2
mutant R177Q
Phanerochaete chrysosporium
1.11.1.13
0.0121
-
H2O2
mutant R177N
Phanerochaete chrysosporium
1.11.1.13
0.0126
-
H2O2
mutant R177D
Phanerochaete chrysosporium
1.11.1.13
0.039
-
H2O2
mutant R177A
Phanerochaete chrysosporium
1.11.1.13
0.044
-
H2O2
mutant R177K
Phanerochaete chrysosporium
1.11.1.13
0.055
-
H2O2
wild-type MnP
Phanerochaete chrysosporium
1.11.1.13
0.09
-
Mn2+
wild-type MnP
Phanerochaete chrysosporium
1.11.1.13
1.64
-
Mn2+
mutant R177A
Phanerochaete chrysosporium
1.11.1.13
2.15
-
Mn2+
mutant R177D
Phanerochaete chrysosporium
1.11.1.13
2.32
-
Mn2+
mutant R177K
Phanerochaete chrysosporium
1.11.1.13
2.91
-
Mn2+
mutant R177Q
Phanerochaete chrysosporium
1.11.1.13
3.61
-
Mn2+
mutant R177N
Phanerochaete chrysosporium
1.11.1.13
3.9
-
Mn2+
mutant R177E
Phanerochaete chrysosporium
1.11.1.13
4.4
-
Mn2+
mutant E35Q
Phanerochaete chrysosporium
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.11.1.13
extracellular
-
Phanerochaete chrysosporium
-
-
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.11.1.13
46000
-
x * 46000, wild-type and mutants R177D, R177E, R177N and R177Q, SDS-PAGE
Phanerochaete chrysosporium
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.11.1.13
Mn2+ + H+ + H2O2
Phanerochaete chrysosporium
important component of lignin degradation system
Mn3+ + H2O
-
Phanerochaete chrysosporium
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.11.1.13
purification of the mutant enzymes R177D, R177E, R177N and R177Q
Phanerochaete chrysosporium
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.11.1.13
Mn2+ + H+ + H2O2
unique binding and oxidation site for Mn2+, single Mn atom is hexacoordinate, with two water ligands and four carboxylate ligands from heme propionate 6 and amino acids Glu-35, Glu-39 and Asp-179
439839
Phanerochaete chrysosporium
Mn3+ + H2O
freely diffusible, enzyme-generated Mn(III)-organic-acid complex oxidizes phenolic substrates
439839
Phanerochaete chrysosporium
-
1.11.1.13
Mn2+ + H+ + H2O2
unique binding and oxidation site for Mn2+, single Mn atom is hexacoordinate, with two water ligands and four carboxylate ligands from heme propionate 6 and amino acids Glu-35, Glu-39 and Asp-179
439839
Phanerochaete chrysosporium
Mn3+ + H2O
Mn3+ oxidizes lignin
439839
Phanerochaete chrysosporium
-
1.11.1.13
Mn2+ + H+ + H2O2
role for Arg-177 in promoting efficient Mn2+ binding and oxidation by MnP
439839
Phanerochaete chrysosporium
Mn3+ + H2O
freely diffusible, enzyme-generated Mn(III)-organic-acid complex oxidizes phenolic substrates
439839
Phanerochaete chrysosporium
-
1.11.1.13
Mn2+ + H+ + H2O2
role for Arg-177 in promoting efficient Mn2+ binding and oxidation by MnP
439839
Phanerochaete chrysosporium
Mn3+ + H2O
Mn3+ oxidizes lignin
439839
Phanerochaete chrysosporium
-
1.11.1.13
Mn2+ + H+ + H2O2
important component of lignin degradation system
439839
Phanerochaete chrysosporium
Mn3+ + H2O
-
439839
Phanerochaete chrysosporium
?
1.11.1.13
Mn2+ + H+ + H2O2
unique binding and oxidation site for Mn2+, single Mn atom is hexacoordinate, with two water ligands and four carboxylate ligands from heme propionate 6 and amino acids Glu-35, Glu-39 and Asp-179
439839
Phanerochaete chrysosporium OGC101
Mn3+ + H2O
freely diffusible, enzyme-generated Mn(III)-organic-acid complex oxidizes phenolic substrates
439839
Phanerochaete chrysosporium OGC101
-
1.11.1.13
additional information
structural properties
439839
Phanerochaete chrysosporium
?
-
-
-
-
1.11.1.13
additional information
catalytic cycle with oxidized intermediates MnP compound I and II
439839
Phanerochaete chrysosporium
?
-
-
-
-
1.11.1.13
additional information
enzyme oxidizes ferrocyanide
439839
Phanerochaete chrysosporium
?
-
-
-
-
1.11.1.13
additional information
enzyme oxidizes bromide
439839
Phanerochaete chrysosporium
?
-
-
-
-
1.11.1.13
additional information
structural properties
439839
Phanerochaete chrysosporium OGC101
?
-
-
-
-
1.11.1.13
additional information
catalytic cycle with oxidized intermediates MnP compound I and II
439839
Phanerochaete chrysosporium OGC101
?
-
-
-
-
1.11.1.13
additional information
enzyme oxidizes ferrocyanide
439839
Phanerochaete chrysosporium OGC101
?
-
-
-
-
1.11.1.13
additional information
enzyme oxidizes bromide
439839
Phanerochaete chrysosporium OGC101
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.11.1.13
?
x * 46000, wild-type and mutants R177D, R177E, R177N and R177Q, SDS-PAGE
Phanerochaete chrysosporium
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.11.1.13
0.77
-
Mn2+
mutant E35Q
Phanerochaete chrysosporium
1.11.1.13
17.3
-
H2O2
mutant R177E
Phanerochaete chrysosporium
1.11.1.13
23.2
-
H2O2
mutant R177Q
Phanerochaete chrysosporium
1.11.1.13
25
-
Mn2+
mutant R177Q
Phanerochaete chrysosporium
1.11.1.13
25.5
-
Mn2+
mutant R177E
Phanerochaete chrysosporium
1.11.1.13
28.8
-
H2O2
mutant R177D
Phanerochaete chrysosporium
1.11.1.13
30.2
-
Mn2+
mutant R177D
Phanerochaete chrysosporium
1.11.1.13
34.6
-
Mn2+
mutant R177N
Phanerochaete chrysosporium
1.11.1.13
35.8
-
H2O2
mutant R177N
Phanerochaete chrysosporium
1.11.1.13
184
-
Mn2+
mutant R177A
Phanerochaete chrysosporium
1.11.1.13
190
-
H2O2
mutant R177A
Phanerochaete chrysosporium
1.11.1.13
252
-
H2O2
wild-type MnP
Phanerochaete chrysosporium
1.11.1.13
256
-
Mn2+
wild-type MnP
Phanerochaete chrysosporium
1.11.1.13
262
-
Mn2+
mutant R177K
Phanerochaete chrysosporium
1.11.1.13
273
-
H2O2
mutant R177K
Phanerochaete chrysosporium