1ALQ

CIRCULARLY PERMUTED BETA-LACTAMASE FROM STAPHYLOCOCCUS AUREUS PC1


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 1.80 Å
  • R-Value Free: 0.256 
  • R-Value Observed: 0.191 

wwPDB Validation   3D Report Full Report


This is version 1.3 of the entry. See complete history


Literature

Circularly permuted beta-lactamase from Staphylococcus aureus PC1.

Pieper, U.Hayakawa, K.Li, Z.Herzberg, O.

(1997) Biochemistry 36: 8767-8774

  • DOI: https://doi.org/10.1021/bi9705117
  • Primary Citation of Related Structures:  
    1ALQ

  • PubMed 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.


  • Organizational Affiliation

    Center for Advanced Research in Biotechnology, University of Maryland Biotechnology Institute, 9600 Gudelsky Drive, Rockville, Maryland 20850, USA.


Macromolecules
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
CP254 BETA-LACTAMASE266Staphylococcus aureusMutation(s): 0 
EC: 3.5.2.6
UniProt
Find proteins for P00807 (Staphylococcus aureus)
Explore P00807 
Go to UniProtKB:  P00807
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupP00807
Sequence Annotations
Expand
  • Reference Sequence
Small Molecules
Ligands 2 Unique
IDChains Name / Formula / InChI Key2D Diagram3D Interactions
SO4
Query on SO4

Download Ideal Coordinates CCD File 
B [auth A]SULFATE ION
O4 S
QAOWNCQODCNURD-UHFFFAOYSA-L
CO3
Query on CO3

Download Ideal Coordinates CCD File 
C [auth A]CARBONATE ION
C O3
BVKZGUZCCUSVTD-UHFFFAOYSA-L
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 1.80 Å
  • R-Value Free: 0.256 
  • R-Value Observed: 0.191 
  • Space Group: I 2 2 2
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 53.9α = 90
b = 94.2β = 90
c = 138.3γ = 90
Software Package:
Software NamePurpose
SHELXL-97model building
X-PLORmodel building
SHELXL-97refinement
X-PLORrefinement
XENGENdata reduction
XENGENdata scaling
SHELXL-97phasing
X-PLORphasing

Structure Validation

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Entry History 

Deposition Data

Revision History  (Full details and data files)

  • Version 1.0: 1997-09-26
    Type: Initial release
  • Version 1.1: 2008-03-24
    Changes: Version format compliance
  • Version 1.2: 2011-07-13
    Changes: Version format compliance
  • Version 1.3: 2023-08-02
    Changes: Database references, Derived calculations, Other, Refinement description