 |
|
|
|
|
|
 |
Contents |
 |
|
|
|
|
|
|
|
|
|
|
|
|
* Residue conservation analysis
|
|
|
|
|
|
|
|
|
|
|
|
Gene Ontology (GO) functional annotation
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Biological process
|
response to antibiotic
|
2 terms
|
|
|
Biochemical function
|
beta-lactamase activity
|
1 term
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
| |
|
DOI no:
|
Biochemistry
36:8767-8774
(1997)
|
|
PubMed id:
|
|
|
|
|
|
| |
|
Circularly permuted beta-lactamase from Staphylococcus aureus PC1.
|
|
U.Pieper,
K.Hayakawa,
Z.Li,
O.Herzberg.
|
|
|
|
|
| |
ABSTRACT
|
|
|
|
| |
|
|
The role that domain flexibility plays in the enzymatic activity of
beta-lactamase from Staphylococcus aureus PC1 was investigated by producing two
circularly permuted molecules. The C- and N-termini of the wild-type enzyme are
adjacent to each other and remote from the active site, which is located between
two domains. The polypeptide chain crosses over from one domain to the other
twice. For the circularly permuted molecules, the termini were joined by an
eight amino acid residue insertion, and new termini were introduced elsewhere.
The first construct, termed cp254, was cleaved in a loop remote from the domain
interface. The crystal structure of cp254 has been determined and refined at 1.8
A resolution, revealing essentially the same structure as that of the native
protein. The activity profile with a representative sample of beta-lactam
antibiotics is also very similar to that of wild-type beta-lactamase. The
termini of the second circularly permuted mutant, cp228, occur within the second
crossover region and therefore may enhance the flexibility of the molecule.
Cp228 beta-lactamase shows a large decrease in enzymatic activity toward the
sample of beta-lactam antibiotics, with catalytic rates that are 0.5-1% of those
of the wild-type enzyme. One exception is the hydrolysis of the third generation
cephalosporin, cefotaxime, which is hydrolyzed by the cp228 enzyme 10-fold
faster than by wild-type beta-lactamase. Cp228 has not been crystallized.
However, the circular dichroism spectra of the two circularly permuted proteins
are very similar, indicating that, by analogy to cp254, cp228 adopts a global
folded state. Thermal denaturation experiments reveal that cp254 is somewhat
less stable than the wild-type enzyme, whereas cp228 is substantially less
stable. Together, the data highlight the profound consequences that introducing
flexibility at the domain interface has on both enzyme activity and protein
stability.
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
 |
 |
|
Literature references that cite this PDB file's key reference
|
|
|
| |
PubMed id
|
|
Reference
|
|
|
|
|
|
Y.Yu,
and
S.Lutz
(2011).
Circular permutation: a different way to engineer enzyme structure and function.
|
| |
Trends Biotechnol, 29,
18-25.
|
|
|
|
|
|
|
Z.Qian,
J.R.Horton,
X.Cheng,
and
S.Lutz
(2009).
Structural redesign of lipase B from Candida antarctica by circular permutation and incremental truncation.
|
| |
J Mol Biol, 393,
191-201.
|
|
|
PDB codes:
|
|
|
|
|
|
|
|
A.Abyzov,
and
V.A.Ilyin
(2007).
A comprehensive analysis of non-sequential alignments between all protein structures.
|
| |
BMC Struct Biol, 7,
78.
|
|
|
|
|
|
|
Z.Qian,
C.J.Fields,
and
S.Lutz
(2007).
Investigating the structural and functional consequences of circular permutation on lipase B from Candida antarctica.
|
| |
Chembiochem, 8,
1989-1996.
|
|
|
|
|
|
|
B.A.Manjasetty,
J.Hennecke,
R.Glockshuber,
and
U.Heinemann
(2004).
Structure of circularly permuted DsbA(Q100T99): preserved global fold and local structural adjustments.
|
| |
Acta Crystallogr D Biol Crystallogr, 60,
304-309.
|
|
|
PDB code:
|
|
|
|
|
|
|
|
T.U.Schwartz,
R.Walczak,
and
G.Blobel
(2004).
Circular permutation as a tool to reduce surface entropy triggers crystallization of the signal recognition particle receptor beta subunit.
|
| |
Protein Sci, 13,
2814-2818.
|
|
|
|
|
|
|
J.M.Spotts,
R.E.Dolmetsch,
and
M.E.Greenberg
(2002).
Time-lapse imaging of a dynamic phosphorylation-dependent protein-protein interaction in mammalian cells.
|
| |
Proc Natl Acad Sci U S A, 99,
15142-15147.
|
|
|
|
|
|
|
P.T.Beernink,
Y.R.Yang,
R.Graf,
D.S.King,
S.S.Shah,
and
H.K.Schachman
(2001).
Random circular permutation leading to chain disruption within and near alpha helices in the catalytic chains of aspartate transcarbamoylase: effects on assembly, stability, and function.
|
| |
Protein Sci, 10,
528-537.
|
|
|
|
|
|
|
X.Ni,
and
H.K.Schachman
(2001).
In vivo assembly of aspartate transcarbamoylase from fragmented and circularly permuted catalytic polypeptide chains.
|
| |
Protein Sci, 10,
519-527.
|
|
|
|
|
|
|
C.J.Tsai,
J.V.Maizel,
and
R.Nussinov
(1999).
Distinguishing between sequential and nonsequentially folded proteins: implications for folding and misfolding.
|
| |
Protein Sci, 8,
1591-1604.
|
|
|
|
|
|
|
V.Chu,
S.Freitag,
I.Le Trong,
R.E.Stenkamp,
and
P.S.Stayton
(1998).
Thermodynamic and structural consequences of flexible loop deletion by circular permutation in the streptavidin-biotin system.
|
| |
Protein Sci, 7,
848-859.
|
|
|
PDB codes:
|
|
|
|
|
|
|
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
