Literature summary for 1.14.15.1 extracted from

Yang, W.; Bell, S.; Wang, H.; Zhou, W.; Bartlam, M.; Wong, L.; Rao, Z.
The structure of CYP101D2 unveils a potential path for substrate entry into the active site (2011), Biochem. J., 433, 85-93.

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Cloned(Commentary)

Cloned (Comment)	Organism
exxpression of N-terminally His-tagged CYP101D2 in Escherichia coli strain BL21(DE3)	Novosphingobium aromaticivorans

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant apoenzyme and camphor-bound enzyme of CYP101D2, hanging drop vapour diffusion method, mixing of 0.001 ml of 50 mg/ml protein in 20 mM Tris-HCl, pH 8.0, and 150 mM KCl, with 0.001 ml of reservoir solution containing 0.1 M Tris/HCl, pH 8.3, 2.1 M ammonium sulfate, and 4% v/v PEG 400, equilibration against 0.2 ml reservoir solution, 18°C, 1 week, for substrate-bound form soaking of crystals in camphor-containing solution, 18°C, 1 week-1 month, X-ray diffraction structure determination and analysis at 2.4 A and 2.2. A resolution, respectively, molecular replacement and structure modeling	Novosphingobium aromaticivorans

Crystallization (Comment)

Organism

purified recombinant apoenzyme and camphor-bound enzyme of CYP101D2, hanging drop vapour diffusion method, mixing of 0.001 ml of 50 mg/ml protein in 20 mM Tris-HCl, pH 8.0, and 150 mM KCl, with 0.001 ml of reservoir solution containing 0.1 M Tris/HCl, pH 8.3, 2.1 M ammonium sulfate, and 4% v/v PEG 400, equilibration against 0.2 ml reservoir solution, 18°C, 1 week, for substrate-bound form soaking of crystals in camphor-containing solution, 18°C, 1 week-1 month, X-ray diffraction structure determination and analysis at 2.4 A and 2.2. A resolution, respectively, molecular replacement and structure modeling

Novosphingobium aromaticivorans

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
(+)-camphor + reduced ferredoxin + O2	Novosphingobium aromaticivorans	-	(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O	-	?
(+)-camphor + reduced ferredoxin + O2	Novosphingobium aromaticivorans DSM 12444	-	(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Novosphingobium aromaticivorans	Q2G8A2	-	-
Novosphingobium aromaticivorans DSM 12444	Q2G8A2	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged CYP101D2 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, gel filtration, and anion exchange chromatography to over 95% purity	Novosphingobium aromaticivorans

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
(+)-camphor + reduced ferredoxin + O2	-	Novosphingobium aromaticivorans	(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O	-	?
(+)-camphor + reduced ferredoxin + O2	substrate of CYP101D1 and CYP101D2	Novosphingobium aromaticivorans	(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O	-	?
(+)-camphor + reduced ferredoxin + O2	-	Novosphingobium aromaticivorans DSM 12444	(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O	-	?
(+)-camphor + reduced ferredoxin + O2	substrate of CYP101D1 and CYP101D2	Novosphingobium aromaticivorans DSM 12444	(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O	-	?

Synonyms

Synonyms	Comment	Organism
CYP101D1	-	Novosphingobium aromaticivorans
CYP101D2	-	Novosphingobium aromaticivorans

Cofactor

Cofactor	Comment	Organism	Structure
Ferredoxin	the CYP101D2 likely ferredoxin-binding site on the proximal face is largely positively charged, similar to that of CYP101D1	Novosphingobium aromaticivorans
heme	-	Novosphingobium aromaticivorans

General Information

General Information	Comment	Organism
additional information	both the apoenzyme and the camphor-bound enzyme of CYP101D2 have open conformations with an access channel. In the active site of the camphor-bound form, the camphor carbonyl interacts with the heme-iron-bound water. The observed open structures may be conformers of the CYP101D2 enzyme that enable the substrate to enter the buried active site via a conformational selection mechanism. Two other potential camphor-binding sites exist: one located in the access channel, flanked by the B/C and F/G loops and the I helix, and the other in a cavity on the surface of the enzyme near the F helix side of the F/G loop. The second and third binding sites may be intermediate locations of substrate entry and translocation into the active site, substrate binding structure and multi-step substrate-binding mechanism, overview	Novosphingobium aromaticivorans