Literature summary for 1.14.11.2 extracted from

Schnicker, N.; Dey, M.
Bacillus anthracis prolyl 4-hydroxylase modifies collagen-like substrates in asymmetric patterns (2016), J. Biol. Chem., 291, 13360-13374 .

Search for more literature for 1.14.11.2

Cloned(Commentary)

Cloned (Comment)	Organism
gene bap4h, recombinant expression in Escherichia coli strain BL21 star DE3 pLysS cells	Bacillus anthracis

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

Inhibitors	Comment	Organism	Structure
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
malonate	active site binding structure analysis	Bacillus anthracis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	required for catalysis, 1:1 ratio of Fe(II) to BaP4H monomer	Bacillus anthracis

Organism

Organism	UniProt	Comment	Textmining
Bacillus anthracis	Q81LZ8	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme from Escherichia coli strain BL21 star DE3 pLysS cells	Bacillus anthracis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
human procollagen-like (P-P-G)10 peptide L-proline + 2-oxoglutarate + O2	4 different peptide sequences, anaerobic conditions	Bacillus anthracis	human procollagen-like (P-P-G)10 peptide trans-4-hydroxy-L-proline + succinate + CO2	-	?
human procollagen-like (P-P-G)5 peptide L-proline + 2-oxoglutarate + O2	5 different peptide sequences, anaerobic conditions	Bacillus anthracis	human procollagen-like (P-P-G)5 peptide trans-4-hydroxy-L-proline + succinate + CO2	-	?
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	Bacillus anthracis	?	-	?

Synonyms

Synonyms	Comment	Organism
BaP4H	-	Bacillus anthracis
P4H	-	Bacillus anthracis
prolyl 4-hydroxylase	-	Bacillus anthracis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
20	-	assay at	Bacillus anthracis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.4	-	assay at	Bacillus anthracis

Cofactor

Cofactor	Comment	Organism	Structure
ascorbate	-	Bacillus anthracis

General Information

General Information	Comment	Organism
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