PDBsum entry 3w9c

Oxidoreductase/electron transport

PDB id: 3w9c

Contents

Protein chains

401 a.a.

106 a.a.

Ligands

HEM

SO4 ×4

GOL

FES

Waters ×35

PDB id:

3w9c

Name:

Oxidoreductase/electron transport

Title:

Crystal structure of the electron transfer complex of cytochrome p450cam with putidaredoxin

Structure:

Camphor 5-monooxygenase. Chain: a. Synonym: cytochrome p450-cam, cytochrome p450cam. Engineered: yes. Mutation: yes. Putidaredoxin. Chain: b. Synonym: pdx. Engineered: yes.

Source:

Pseudomonas putida. Organism_taxid: 303. Gene: camc, cyp101. Expressed in: escherichia coli. Expression_system_taxid: 562. Gene: camb.

Resolution:

2.50Å R-factor: 0.188 R-free: 0.251

Authors:

Y.Kikui,Y.Hiruma,M.A.Hass,H.Koteishi,M.Ubbink,M.Nojiri

Key ref:

Y.Hiruma et al. (2013). The structure of the cytochrome p450cam-putidaredoxin complex determined by paramagnetic NMR spectroscopy and crystallography. J Mol Biol, 425, 4353-4365. PubMed id: 23856620 DOI: 10.1016/j.jmb.2013.07.006

Date:

03-Apr-13 Release date: 21-Aug-13

Protein chain

P00183  (CPXA_PSEPU) -  Camphor 5-monooxygenase from Pseudomonas putida

Seq: 415 a.a.

Struc: 401 a.a.*

Protein chain

P00259  (PUTX_PSEPU) -  Putidaredoxin from Pseudomonas putida

Seq: 107 a.a.

Struc: 106 a.a.*

* PDB and UniProt seqs differ at 4 residue positions (black crosses)

Enzyme reactions

• Enzyme class: Chain A: E.C.1.14.15.1 - camphor 5-monooxygenase.
• Reaction: 2 reduced [2Fe-2S]-[putidaredoxin] + (1R,4R)-camphor + O2 + 2 H⁺ = (1R,4R,5R)-5-hydroxycamphor + 2 oxidized [2Fe-2S]-[putidaredoxin] + H2O
• Cofactor: Heme-thiolate

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

Key reference

DOI no: 10.1016/j.jmb.2013.07.006

J Mol Biol 425:4353-4365 (2013)

PubMed id: 23856620

The structure of the cytochrome p450cam-putidaredoxin complex determined by paramagnetic NMR spectroscopy and crystallography.

Y.Hiruma, M.A.Hass, Y.Kikui, W.M.Liu, B.Ölmez, S.P.Skinner, A.Blok, A.Kloosterman, H.Koteishi, F.Löhr, H.Schwalbe, M.Nojiri, M.Ubbink.

ABSTRACT

Cytochrome P450cam catalyzes the hydroxylation of camphor in a complex process involving two electron transfers (ETs) from the iron-sulfur protein putidaredoxin. The enzymatic control of the successive steps of catalysis is critical for a highly efficient reaction. The injection of the successive electrons is part of the control system. To understand the molecular interactions between putidaredoxin and cytochrome P450cam, we determined the structure of the complex both in solution and in the crystal state. Paramagnetic NMR spectroscopy using lanthanide tags yielded 446 structural restraints that were used to determine the solution structure. An ensemble of 10 structures with an RMSD of 1.3Å was obtained. The crystal structure of the complex was solved, showing a position of putidaredoxin that is identical with the one in the solution structure. The NMR data further demonstrate the presence of a minor state or set of states of the complex in solution, which is attributed to the presence of an encounter complex. The structure of the major state shows a small binding interface and a metal-to-metal distance of 16Å, with two pathways that provide strong electronic coupling of the redox centers. The interpretation of these results is discussed in the context of ET. The structure indicates that the ET rate can be much faster than the reported value, suggesting that the process may be gated.