Literature summary for 1.14.15.1 extracted from

Nagano, S.; Poulos, T.L.
Crystallographic study on the dioxygen complex of wild-type and mutant cytochrome P450cam. Implications for the dioxygen activation mechanism (2005), J. Biol. Chem., 280, 31659-31663.

Search for more literature for 1.14.15.1

Cloned(Commentary)

Cloned (Comment)	Organism
overexpression of wild-type and mutant enzyme sin Escherichia coli	Pseudomonas putida

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant wild-type and D251N and T252A mutant enzymes in complex with O2, usage of a high pressure oxygen cell, a single crystal first is transferred into cryobuffer containing 50 mM Tris-HCl, pH 7.4, 0.4-0.6 M KCl, 1 mM D-camphor, 30% polyethylene glycol 4000, and 20% glycerol followed by reduction with 10 mM sodium dithionite for 10 min under anaerobic condition, soaking in the oxygen-saturated cryobuffer at -5°C for 5 min, X-ray diffraction structure determination and analysis at 1.55-2.10 A resolution	Pseudomonas putida

Crystallization (Comment)

Organism

purified recombinant wild-type and D251N and T252A mutant enzymes in complex with O2, usage of a high pressure oxygen cell, a single crystal first is transferred into cryobuffer containing 50 mM Tris-HCl, pH 7.4, 0.4-0.6 M KCl, 1 mM D-camphor, 30% polyethylene glycol 4000, and 20% glycerol followed by reduction with 10 mM sodium dithionite for 10 min under anaerobic condition, soaking in the oxygen-saturated cryobuffer at -5°C for 5 min, X-ray diffraction structure determination and analysis at 1.55-2.10 A resolution

Pseudomonas putida

Protein Variants

Protein Variants	Comment	Organism
D251N	site-directed mutagenesis, the mutant shows altered conformation of the I helix groove and misses the catalytically important water molecules in the dioxygen complex leading to lower catalytic activity and slower proton transfer to the dioxygen ligand compared to the wild-type enzyme	Pseudomonas putida
T252A	site-directed mutagenesis, the mutant does not show altered conformation of the I helix groove and the catalytically important water molecules in the dioxygen complex	Pseudomonas putida

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Iron	heme iron	Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
(+)-camphor + O2 + reduced putidaredoxin	Pseudomonas putida	-	(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas putida	-	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
(+)-camphor + O2 + reduced putidaredoxin	-	Pseudomonas putida	(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O	-	?
(+)-camphor + O2 + reduced putidaredoxin	wild-type dioxygen complex structure: high occupancy and a ordered structure of the iron-linked dioxygen and two 'catalytic' water molecules that form part of a proton relay system to the iron-linked dioxygen, Thr252 accepts a hydrogen bond from the hydroperoxy (Fe(III)-OOH) intermediate that promotes the second protonation on the distal oxygen atom, leading to O-O bond cleavage and compound I formation	Pseudomonas putida	(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O	-	?

Subunits

Subunits	Comment	Organism
More	wild-type dioxygen complex structure: high occupancy and a ordered structure of the iron-linked dioxygen and two 'catalytic' water molecules that form part of a proton relay system to the iron-linked dioxygen	Pseudomonas putida

Synonyms

Synonyms	Comment	Organism
cytochrome p450cam	-	Pseudomonas putida

Cofactor

Cofactor	Comment	Organism	Structure
putidaredoxin	-	Pseudomonas putida