 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Oxidoreductase
|
PDB id
|
|
|
|
1iwk
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
Contents |
 |
|
|
|
|
|
|
|
|
|
|
|
|
* Residue conservation analysis
|
|
|
|
|
|
PDB id:
|
|
|
|
| Name: |
|
Oxidoreductase
|
|
|
Title:
|
|
Putidaredoxin-binding stablilizes an active conformer of cytochrome p450cam in its reduced state; crystal structure of mutant(112k) cytochrome p450cam
|
|
Structure:
|
|
Cytochrome p450-cam. Chain: a. Engineered: yes. Mutation: yes
|
|
Source:
|
|
Pseudomonas putida. Organism_taxid: 303. Expressed in: escherichia coli. Expression_system_taxid: 562.
|
|
Resolution:
|
|
|
2.00Å
|
R-factor:
|
0.181
|
R-free:
|
0.235
|
|
|
Authors:
|
|
S.Nagano,H.Shimada,A.Tarumi,T.Hishiki,Y.Kimata-Ariga, T.Egawa,S.-Y.Park,S.Adachi,Y.Shiro,Y.Ishimura,Riken Structural Genomics/proteomics Initiative (Rsgi)
|
Key ref:
|
|
S.Nagano
et al.
(2003).
Infrared spectroscopic and mutational studies on putidaredoxin-induced conformational changes in ferrous CO-P450cam.
Biochemistry,
42,
14507-14514.
PubMed id:
DOI:
|
|
|
Date:
|
|
|
15-May-02
|
Release date:
|
05-Jun-02
|
|
|
|
|
|
|
PROCHECK
|
|
|
|
|
|
Headers
|
|
|
|
References
|
|
|
|
|
|
|
|
P00183
(CPXA_PSEPU) -
Camphor 5-monooxygenase
|
|
|
|
Seq:
Struc:
|
|
|
|
415 a.a.
405 a.a.*
|
|
|
|
|
|
|
|
|
|
|
|
|
Key: |
 |
PfamA domain |
|
|
 |
Secondary structure |
|
 |
CATH domain |
|
|
*
PDB and UniProt seqs differ
at 1 residue position (black
cross)
|
|
|
|
|
|
|
|
|
|
|
|
|
Enzyme class:
|
|
E.C.1.14.15.1
- Camphor 5-monooxygenase.
|
|
|
|
|
|
|
Reaction:
|
|
+-camphor + putidaredoxin + O2 = +-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
|
|
|
|
|
|
(+)-camphor
|
+
|
putidaredoxin
|
+
|
O(2)
|
=
|
(+)-exo-5-hydroxycamphor
|
+
|
oxidized putidaredoxin
|
+
|
H(2)O
|
|
|
|
|
|
|
|
|
|
Cofactor:
|
|
Heme-thiolate
|
|
|
|
|
|
|
|
|
Molecule diagrams generated from .mol files obtained from the
KEGG ftp site
|
|
|
|
|
|
|
|
|
|
|
Gene Ontology (GO) functional annotation
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Cellular component
|
cytoplasm
|
1 term
|
|
|
Biological process
|
oxidation reduction
|
1 term
|
|
|
Biochemical function
|
electron carrier activity
|
7 terms
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
| |
|
|
| |
|
DOI no:
|
Biochemistry
42:14507-14514
(2003)
|
|
PubMed id:
|
|
|
|
|
|
| |
|
Infrared spectroscopic and mutational studies on putidaredoxin-induced conformational changes in ferrous CO-P450cam.
|
|
S.Nagano,
H.Shimada,
A.Tarumi,
T.Hishiki,
Y.Kimata-Ariga,
T.Egawa,
M.Suematsu,
S.Y.Park,
S.Adachi,
Y.Shiro,
Y.Ishimura.
|
|
|
|
|
| |
ABSTRACT
|
|
|
|
| |
|
|
Ferrous-carbon monoxide bound form of cytochrome P450cam (CO-P450cam) has two
infrared (IR) CO stretching bands at 1940 and 1932 cm(-1). The former band is
dominant (>95% in area) for CO-P450cam free of putidaredoxin (Pdx), while the
latter band is dominant (>95% in area) in the complex of CO-P450cam with reduced
Pdx. The binding of Pdx to CO-P450cam thus evokes a conformational change in the
heme active site. To study the mechanism involved in the conformational change,
surface amino acid residues Arg79, Arg109, and Arg112 in P450cam were replaced
with Lys, Gln, and Met. IR spectroscopic and kinetic analyses of the mutants
revealed that an enzyme that has a larger 1932 cm(-1) band area upon Pdx-binding
has a larger catalytic activity. Examination of the crystal structures of R109K
and R112K suggested that the interaction between the guanidium group of Arg112
and Pdx is important for the conformational change. The mutations did not change
a coupling ratio between the hydroxylation product and oxygen consumed. We
interpret these findings to mean that the interaction of P450cam with Pdx
through Arg112 enhances electron donation from the proximal ligand (Cys357) to
the O-O bond of iron-bound O(2) and, possibly, promotes electron transfer from
reduced Pdx to oxyP450cam, thereby facilitating the O-O bond splitting.
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
 |
 |
|
Literature references that cite this PDB file's key reference
|
|
|
| |
PubMed id
|
|
Reference
|
|
|
|
|
|
H.Kabumoto,
K.Miyazaki,
and
A.Arisawa
(2009).
Directed evolution of the actinomycete cytochrome P450moxA (CYP105) for enhanced activity.
|
| |
Biosci Biotechnol Biochem, 73,
1922-1927.
|
|
|
|
|
|
|
C.Jung
(2008).
Fourier transform infrared spectroscopy as a tool to study structural properties of cytochromes P450 (CYPs).
|
| |
Anal Bioanal Chem, 392,
1031-1058.
|
|
|
|
|
|
|
T.Tosha,
N.Kagawa,
M.Arase,
M.R.Waterman,
and
T.Kitagawa
(2008).
Interaction between substrate and oxygen ligand responsible for effective O-O bond cleavage in bovine cytochrome P450 steroid 21-hydroxylase proved by Raman spectroscopy.
|
| |
J Biol Chem, 283,
3708-3717.
|
|
|
|
|
|
|
M.Ibrahim,
C.Xu,
and
T.G.Spiro
(2006).
Differential sensing of protein influences by NO and CO vibrations in heme adducts.
|
| |
J Am Chem Soc, 128,
16834-16845.
|
|
|
|
|
|
|
V.Y.Kuznetsov,
T.L.Poulos,
and
I.F.Sevrioukova
(2006).
Putidaredoxin-to-cytochrome P450cam electron transfer: differences between the two reductive steps required for catalysis.
|
| |
Biochemistry, 45,
11934-11944.
|
|
|
|
|
|
|
M.C.Glascock,
D.P.Ballou,
and
J.H.Dawson
(2005).
Direct observation of a novel perturbed oxyferrous catalytic intermediate during reduced putidaredoxin-initiated turnover of cytochrome P-450-CAM: probing the effector role of putidaredoxin in catalysis.
|
| |
J Biol Chem, 280,
42134-42141.
|
|
|
|
|
|
|
K.Konishi,
K.Ishida,
K.Oinuma,
T.Ohta,
Y.Hashimoto,
H.Higashibata,
T.Kitagawa,
and
M.Kobayashi
(2004).
Identification of crucial histidines involved in carbon-nitrogen triple bond synthesis by aldoxime dehydratase.
|
| |
J Biol Chem, 279,
47619-47625.
|
|
|
|
|
|
|
S.Nagano,
T.Tosha,
K.Ishimori,
I.Morishima,
and
T.L.Poulos
(2004).
Crystal structure of the cytochrome p450cam mutant that exhibits the same spectral perturbations induced by putidaredoxin binding.
|
| |
J Biol Chem, 279,
42844-42849.
|
|
|
PDB codes:
|
|
|
|
|
|
|
|
T.Tosha,
S.Yoshioka,
K.Ishimori,
and
I.Morishima
(2004).
L358P mutation on cytochrome P450cam simulates structural changes upon putidaredoxin binding: the structural changes trigger electron transfer to oxy-P450cam from electron donors.
|
| |
J Biol Chem, 279,
42836-42843.
|
|
|
|
|
|
The most recent references are shown first.
Citation data come partly from CiteXplore and partly
from an automated harvesting procedure. Note that this is likely to be
only a partial list as not all journals are covered by
either method. However, we are continually building up the citation data
so more and more references will be included with time.
Where a reference describes a PDB structure, the PDB
codes are
shown on the right.
|
|
Links
