 |
PDBsum entry 1cyf
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Oxidoreductase (h2o2(a))
|
PDB id
|
|
|
|
1cyf
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
References listed in PDB file
|
|
|
Key reference
|
|
|
Title
|
|
Identifying the physiological electron transfer site of cytochrome c peroxidase by structure-Based engineering.
|
|
|
Authors
|
|
M.A.Miller,
L.Geren,
G.W.Han,
A.Saunders,
J.Beasley,
G.J.Pielak,
B.Durham,
F.Millett,
J.Kraut.
|
|
|
Ref.
|
|
Biochemistry, 1996,
35,
667-673.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
Note In the PDB file this reference is
annotated as "TO BE PUBLISHED".
The citation details given above were identified by an automated
search of PubMed on title and author
names, giving a
percentage match of
83%.
|
|
|
|
Abstract
|
|
|
A technique was developed to evaluate whether electron transfer (ET) complexes
formed in solution by the cloned cytochrome c peroxidase [CcP(MI)] and
cytochromes c from yeast (yCc) and horse (hCc) are structurally similar to those
seen in the respective crystal structures. Site-directed mutagenesis was used to
convert the sole Cys of the parent enzyme (Cys 128) to Ala, and a Cys residue
was introduced at position 193 of CcP(MI), the point of closest contact between
CcP(MI) and yCc in the crystal structure. Cys 193 was then modified with a bulky
sulfhydryl reagent, 3-(N-maleimidylpropionyl)-biocytin (MPB), to prevent yCc
from binding at the site seen in the crystal. The MPB modification has no effect
on overall enzyme structure but causes 20-100-fold decreases in transient and
steady-state ET reaction rates with yCc. The MPB modification causes only
2-3-fold decreases in ET reaction rates with hCc, however. This differential
effect is predicted by modeling studies based on the crystal structures and
indicates that solution phase ET complexes closely resemble the crystalline
complexes. The low rate of catalysis of the MPB-enzyme was constant for yCc in
buffers of 20-160 mM ionic strength. This indicates that the low affinity
complex formed between CcP(MI) and yCc at low ionic strength is not reactive in
ET.
|
|
|
Secondary reference #1
|
|
|
Title
|
|
X-Ray structures of recombinant yeast cytochrome c peroxidase and three heme-Cleft mutants prepared by site-Directed mutagenesis.
|
|
|
Authors
|
|
J.M.Wang,
M.Mauro,
S.L.Edwards,
S.J.Oatley,
L.A.Fishel,
V.A.Ashford,
N.H.Xuong,
J.Kraut.
|
|
|
Ref.
|
|
Biochemistry, 1990,
29,
7160-7173.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
|
|
|
|
Secondary reference #2
|
|
|
Title
|
|
Crystal structure of yeast cytochrome c peroxidase refined at 1.7-A resolution.
|
|
|
Authors
|
|
B.C.Finzel,
T.L.Poulos,
J.Kraut.
|
|
|
Ref.
|
|
J Biol Chem, 1984,
259,
13027-13036.
|
|
|
PubMed id
|
|
|
|
|
|
|
|
|
|
|
|
|
