PDBsum entry 3awm

Oxidoreductase

PDB id: 3awm

Contents

Protein chain

407 a.a.

Ligands

HEM

PLM

MRD

Waters ×368

PDB id:

3awm

Name:

Oxidoreductase

Title:

Cytochrome p450sp alpha (cyp152b1) wild-type with palmitic acid

Structure:

Fatty acid alpha-hydroxylase. Chain: a. Synonym: cytochrome p450sp alpha, cyp152b1. Engineered: yes

Source:

Sphingomonas paucimobilis. Organism_taxid: 13689. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.65Å R-factor: 0.152 R-free: 0.173

Authors:

T.Fujishiro,O.Shoji,S.Nagano,H.Sugimoto,Y.Shiro,Y.Watanabe

Key ref:

T.Fujishiro et al. (2011). Crystal structure of H2O2-dependent cytochrome P450SPalpha with its bound fatty acid substrate: insight into the regioselective hydroxylation of fatty acids at the alpha position. J Biol Chem, 286, 29941-29950. PubMed id: 21719702

Date:

25-Mar-11 Release date: 29-Jun-11

Protein chain

O24782  (O24782_SPHPI) -  Cytochrome P450 from Sphingomonas paucimobilis

Seq: 415 a.a.

Struc: 407 a.a.

Enzyme reactions

• Enzyme class: E.C.1.11.2.4 - fatty-acid peroxygenase.
• Reaction:
  1. a 1,2-saturated fatty acid + H2O2 = a 2-hydroxy fatty acid + H2O
  2. a 2,3-saturated fatty acid + H2O2 = a 3-hydroxy fatty acid + H2O
• Cofactor: Heme-thiolate

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Added reference

J Biol Chem 286:29941-29950 (2011)

PubMed id: 21719702

Crystal structure of H2O2-dependent cytochrome P450SPalpha with its bound fatty acid substrate: insight into the regioselective hydroxylation of fatty acids at the alpha position.

T.Fujishiro, O.Shoji, S.Nagano, H.Sugimoto, Y.Shiro, Y.Watanabe.

ABSTRACT

Cytochrome P450(SPα) (CYP152B1) isolated from Sphingomonas paucimobilis is the first P450 to be classified as a H(2)O(2)-dependent P450. P450(SPα) hydroxylates fatty acids with high α-regioselectivity. Herein we report the crystal structure of P450(SPα) with palmitic acid as a substrate at a resolution of 1.65 Å. The structure revealed that the C(α) of the bound palmitic acid in one of the alternative conformations is 4.5 Å from the heme iron. This conformation explains the highly selective α-hydroxylation of fatty acid observed in P450(SPα). Mutations at the active site and the F-G loop of P450(SPα) did not impair its regioselectivity. The crystal structures of mutants (L78F and F288G) revealed that the location of the bound palmitic acid was essentially the same as that in the WT, although amino acids at the active site were replaced with the corresponding amino acids of cytochrome P450(BSβ) (CYP152A1), which shows β-regioselectivity. This implies that the high regioselectivity of P450(SPα) is caused by the orientation of the hydrophobic channel, which is more perpendicular to the heme plane than that of P450(BSβ).