PDBsum entry 1alq

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Hydrolase PDB id
Protein chain
259 a.a. *
Waters ×212
* Residue conservation analysis
PDB id:
Name: Hydrolase
Title: Circularly permuted beta-lactamase from staphylococcus aureus pc1
Structure: Cp254 beta-lactamase. Chain: a. Engineered: yes. Mutation: yes
Source: Staphylococcus aureus. Organism_taxid: 1280. Cell_line: 293. Expressed in: escherichia coli. Expression_system_taxid: 562.
1.80Å     R-factor:   0.196     R-free:   0.256
Authors: U.Pieper,O.Herzberg
Key ref:
U.Pieper et al. (1997). Circularly permuted beta-lactamase from Staphylococcus aureus PC1. Biochemistry, 36, 8767-8774. PubMed id: 9220963 DOI: 10.1021/bi9705117
02-Jun-97     Release date:   26-Sep-97    
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Protein chain
Pfam   ArchSchema ?
P00807  (BLAC_STAAU) -  Beta-lactamase
281 a.a.
259 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 21 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: E.C.  - Beta-lactamase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]

Penicillin Biosynthesis and Metabolism
      Reaction: A beta-lactam + H2O = a substituted beta-amino acid
      Cofactor: Zn(2+)
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     response to antibiotic   2 terms 
  Biochemical function     beta-lactamase activity     1 term  


DOI no: 10.1021/bi9705117 Biochemistry 36:8767-8774 (1997)
PubMed id: 9220963  
Circularly permuted beta-lactamase from Staphylococcus aureus PC1.
U.Pieper, K.Hayakawa, Z.Li, O.Herzberg.
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
21087800 Y.Yu, and S.Lutz (2011).
Circular permutation: a different way to engineer enzyme structure and function.
  Trends Biotechnol, 29, 18-25.  
19683009 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: 3icv 3icw
18005453 A.Abyzov, and V.A.Ilyin (2007).
A comprehensive analysis of non-sequential alignments between all protein structures.
  BMC Struct Biol, 7, 78.  
17876754 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.  
14747707 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: 1un2
15340174 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.  
12415118 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.  
11344321 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.  
11344320 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.  
  10452603 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.  
  9568892 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: 1swf 1swg
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