BRENDA - Enzyme Database

Bacillus anthracis prolyl 4-hydroxylase modifies collagen-like substrates in asymmetric patterns

Schnicker, N.; Dey, M.; J. Biol. Chem. 291, 13360-13374 (2016)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.14.11.2
gene bap4h, recombinant expression in Escherichia coli strain BL21 star DE3 pLysS cells
Bacillus anthracis
Crystallization (Commentary)
EC Number
Crystallization
Organism
1.14.11.2
purified recombinant apoenzyme in complex with inhibitor malonate and Co2+, hanging drop vapor diffusion, mixing of 0.001 ml of a solution of 9 mg/ml protein, 1 mM CoCl2, and 1 mM L-Pro, with 0.001 ml of reservoir solution containing 0.1 M malonate/imidazole/boric acid, pH 6.5, and 18% PEG 1500, 2 days, or purified recombinant apoenzyme BaP4H with (P-P-G)3 peptide fused to the C terminus and Co2+, sitting drop vapor diffusion, mixing of 0.001 ml of 12 mg/ml protein solution containing 1 mM CoCl2, and 1 mM 2-oxoglutarate, with 0.001 ml of reservoir solution containing 0.15 M KBr and 30% PEG 2000 MME, 20°C, method optimization, X-ray diffraction structure determination and analysis, molecular replacement for apo-BaP4H using structure PDB ID 3ITQ as a starting model. Models for Co(II)-BaP4H-MLI and Co(II)-BaP4H-PPG are obtained by molecular replacement with the apo-BaP4H structure
Bacillus anthracis
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.14.11.2
Co2+
active site binding structure analysis, in both Co(II)-bound structures, in addition to the catalytic triad residues and 2-oxoglutarate or, malonate that coordinate cobalt, the remaining coordinate sites are occupied by water molecules thus forming a six-coordinate cobalt complex
Bacillus anthracis
1.14.11.2
malonate
active site binding structure analysis
Bacillus anthracis
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.14.11.2
Fe2+
required for catalysis, 1:1 ratio of Fe(II) to BaP4H monomer
Bacillus anthracis
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.14.11.2
Bacillus anthracis
Q81LZ8
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.14.11.2
recombinant enzyme from Escherichia coli strain BL21 star DE3 pLysS cells
Bacillus anthracis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.11.2
human procollagen-like (P-P-G)10 peptide L-proline + 2-oxoglutarate + O2
4 different peptide sequences, anaerobic conditions
745342
Bacillus anthracis
human procollagen-like (P-P-G)10 peptide trans-4-hydroxy-L-proline + succinate + CO2
-
-
-
?
1.14.11.2
human procollagen-like (P-P-G)5 peptide L-proline + 2-oxoglutarate + O2
5 different peptide sequences, anaerobic conditions
745342
Bacillus anthracis
human procollagen-like (P-P-G)5 peptide trans-4-hydroxy-L-proline + succinate + CO2
-
-
-
?
1.14.11.2
additional information
enzyme BaP4H recognizes and acts on peptidyl substrates but not free L-proline, using elements characteristic of an Fe(II)/2-oxoglutarate-dependent dioxygenases, substrate specifiicty analysis by mass spectrometry, fluorescence binding, x-ray crystallography, and docking experiments, overview. Enzyme BaP4H can hydroxylate unique peptidyl proline sites in collagen-derived peptides with asymmetric hydroxylation patterns. The cofactor-bound crystal structures of BaP4H reveal active site conformational changes that define open and closed forms and mimic 'ready' and 'product-released' states of the enzyme in the catalytic cycle. Free L-proline binds to Fe(II)-BaP4H, but the affinity is an order of magnitude lower than for the peptides. No activity with free L-proline
745342
Bacillus anthracis
?
-
-
-
-
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.14.11.2
20
-
assay at
Bacillus anthracis
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.14.11.2
7.4
-
assay at
Bacillus anthracis
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.14.11.2
ascorbate
-
Bacillus anthracis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.11.2
gene bap4h, recombinant expression in Escherichia coli strain BL21 star DE3 pLysS cells
Bacillus anthracis
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.14.11.2
ascorbate
-
Bacillus anthracis
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.14.11.2
purified recombinant apoenzyme in complex with inhibitor malonate and Co2+, hanging drop vapor diffusion, mixing of 0.001 ml of a solution of 9 mg/ml protein, 1 mM CoCl2, and 1 mM L-Pro, with 0.001 ml of reservoir solution containing 0.1 M malonate/imidazole/boric acid, pH 6.5, and 18% PEG 1500, 2 days, or purified recombinant apoenzyme BaP4H with (P-P-G)3 peptide fused to the C terminus and Co2+, sitting drop vapor diffusion, mixing of 0.001 ml of 12 mg/ml protein solution containing 1 mM CoCl2, and 1 mM 2-oxoglutarate, with 0.001 ml of reservoir solution containing 0.15 M KBr and 30% PEG 2000 MME, 20°C, method optimization, X-ray diffraction structure determination and analysis, molecular replacement for apo-BaP4H using structure PDB ID 3ITQ as a starting model. Models for Co(II)-BaP4H-MLI and Co(II)-BaP4H-PPG are obtained by molecular replacement with the apo-BaP4H structure
Bacillus anthracis
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.14.11.2
Co2+
active site binding structure analysis, in both Co(II)-bound structures, in addition to the catalytic triad residues and 2-oxoglutarate or, malonate that coordinate cobalt, the remaining coordinate sites are occupied by water molecules thus forming a six-coordinate cobalt complex
Bacillus anthracis
1.14.11.2
malonate
active site binding structure analysis
Bacillus anthracis
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.14.11.2
Fe2+
required for catalysis, 1:1 ratio of Fe(II) to BaP4H monomer
Bacillus anthracis
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.11.2
recombinant enzyme from Escherichia coli strain BL21 star DE3 pLysS cells
Bacillus anthracis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.11.2
human procollagen-like (P-P-G)10 peptide L-proline + 2-oxoglutarate + O2
4 different peptide sequences, anaerobic conditions
745342
Bacillus anthracis
human procollagen-like (P-P-G)10 peptide trans-4-hydroxy-L-proline + succinate + CO2
-
-
-
?
1.14.11.2
human procollagen-like (P-P-G)5 peptide L-proline + 2-oxoglutarate + O2
5 different peptide sequences, anaerobic conditions
745342
Bacillus anthracis
human procollagen-like (P-P-G)5 peptide trans-4-hydroxy-L-proline + succinate + CO2
-
-
-
?
1.14.11.2
additional information
enzyme BaP4H recognizes and acts on peptidyl substrates but not free L-proline, using elements characteristic of an Fe(II)/2-oxoglutarate-dependent dioxygenases, substrate specifiicty analysis by mass spectrometry, fluorescence binding, x-ray crystallography, and docking experiments, overview. Enzyme BaP4H can hydroxylate unique peptidyl proline sites in collagen-derived peptides with asymmetric hydroxylation patterns. The cofactor-bound crystal structures of BaP4H reveal active site conformational changes that define open and closed forms and mimic 'ready' and 'product-released' states of the enzyme in the catalytic cycle. Free L-proline binds to Fe(II)-BaP4H, but the affinity is an order of magnitude lower than for the peptides. No activity with free L-proline
745342
Bacillus anthracis
?
-
-
-
-
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.14.11.2
20
-
assay at
Bacillus anthracis
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.14.11.2
7.4
-
assay at
Bacillus anthracis
General Information
EC Number
General Information
Commentary
Organism
1.14.11.2
additional information
ligand docking calculations, substrate and inhibitor binding structure, structure-function analysis, coformations of key active site residues Tyr118, Tyr124, and Phe160 in apoenzyme and ligand-bound enzyme, conformational changes involving residues Tyr118, Tyr124, and Phe160 upon metal and 2-oxoglutarate or malonate binding, detailed overview
Bacillus anthracis
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.14.11.2
additional information
ligand docking calculations, substrate and inhibitor binding structure, structure-function analysis, coformations of key active site residues Tyr118, Tyr124, and Phe160 in apoenzyme and ligand-bound enzyme, conformational changes involving residues Tyr118, Tyr124, and Phe160 upon metal and 2-oxoglutarate or malonate binding, detailed overview
Bacillus anthracis