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PDBsum entry 2rl3
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Contents |
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* Residue conservation analysis
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PDB id:
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Hydrolase
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Title:
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Crystal structure of the oxa-10 w154h mutant at ph 7
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Structure:
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Beta-lactamase pse-2. Chain: a, b. Synonym: beta lactamase oxa-10. Engineered: yes. Mutation: yes
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Source:
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Pseudomonas aeruginosa. Organism_taxid: 287. Gene: pse2, oxa10. Expressed in: escherichia coli. Expression_system_taxid: 562.
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Resolution:
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1.90Å
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R-factor:
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0.171
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R-free:
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0.218
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Authors:
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L.Vercheval,F.Kerff,R.Herman,E.Sauvage,R.Guiet,P.Charlier,J.-M.Frere, M.Galleni
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Key ref:
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S.Baurin
et al.
(2009).
Critical role of tryptophan 154 for the activity and stability of class D beta-lactamases.
Biochemistry,
48,
11252-11263.
PubMed id:
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Date:
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18-Oct-07
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Release date:
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28-Oct-08
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PROCHECK
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Headers
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References
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P14489
(BLO10_PSEAI) -
Beta-lactamase OXA-10 from Pseudomonas aeruginosa
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Seq:
Struc:
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266 a.a.
247 a.a.*
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Key: |
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PfamA domain |
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Secondary structure |
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CATH domain |
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*
PDB and UniProt seqs differ
at 3 residue positions (black
crosses)
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Biochemistry
48:11252-11263
(2009)
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PubMed id:
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Critical role of tryptophan 154 for the activity and stability of class D beta-lactamases.
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S.Baurin,
L.Vercheval,
F.Bouillenne,
C.Falzone,
A.Brans,
L.Jacquamet,
J.L.Ferrer,
E.Sauvage,
D.Dehareng,
J.M.Frère,
P.Charlier,
M.Galleni,
F.Kerff.
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ABSTRACT
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The catalytic efficiency of the class D beta-lactamase OXA-10 depends critically
on an unusual carboxylated lysine as the general base residue for both the
enzyme acylation and deacylation steps of catalysis. Evidence is presented that
the interaction between the indole group of Trp154 and the carboxylated lysine
is essential for the stability of the posttranslationally modified Lys70.
Substitution of Trp154 by Gly, Ala, or Phe yielded noncarboxylated enzymes which
displayed poor catalytic efficiencies and reduced stability when compared to the
wild-type OXA-10. The W154H mutant was partially carboxylated. In addition, the
maximum values of k(cat) and k(cat)/K(M) were shifted toward pH 7, indicating
that the carboxylation state of Lys70 is dependent on the protonation level of
the histidine. A comparison of the three-dimensional structures of the different
proteins also indicated that the Trp154 mutations did not modify the overall
structures of OXA-10 but induced an increased flexibility of the Omega-loop in
the active site. Finally, the deacylation-impaired W154A mutant was used to
determine the structure of the acyl-enzyme complex with benzylpenicillin. These
results indicate a role of the Lys70 carboxylation during the deacylation step
and emphasize the importance of Trp154 for the ideal positioning of active site
residues leading to an optimum activity.
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Literature references that cite this PDB file's key reference
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PubMed id
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Reference
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L.Vercheval,
C.Bauvois,
A.di Paolo,
F.Borel,
J.L.Ferrer,
E.Sauvage,
A.Matagne,
J.M.Frère,
P.Charlier,
M.Galleni,
and
F.Kerff
(2010).
Three factors that modulate the activity of class D β-lactamases and interfere with the post-translational carboxylation of Lys70.
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Biochem J,
432,
495-504.
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PDB codes:
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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.
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Links
