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PDBsum entry 2wkj

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protein ligands Protein-protein interface(s) links
Lyase PDB id
2wkj
Jmol
Contents
Protein chains
298 a.a. *
Ligands
1PE ×2
PYR ×2
Waters ×1307
* Residue conservation analysis
PDB id:
2wkj
Name: Lyase
Title: Crystal structure of the e192n mutant of e.Coli n-acetylneuraminic acid lyase in complex with pyruvate at 1.45a resolution in space group p212121
Structure: N-acetylneuraminate lyase. Chain: a, b, c, d. Fragment: residues 2-296. Synonym: n-acetylneuraminic acid, n-acetylneuraminic acid a n-acetylneuraminate pyruvate-lyase, sialic acid lyase, sia lyase, sialic acid aldolase, nalase. Engineered: yes. Mutation: yes. Other_details: schiff base between k165 and pyruvate
Source: Escherichia coli. Organism_taxid: 562. Expressed in: escherichia coli. Expression_system_taxid: 562.
Resolution:
1.45Å     R-factor:   0.167     R-free:   0.188
Authors: I.Campeotto,S.B.Carr,C.H.Trinh,A.S.Nelson,A.Berry,S.E.V.Phil A.R.Pearson
Key ref: I.Campeotto et al. (2009). Structure of an Escherichia coli N-acetyl-D-neuraminic acid lyase mutant, E192N, in complex with pyruvate at 1.45 angstrom resolution. Acta Crystallogr Sect F Struct Biol Cryst Commun, 65, 1088-1090. PubMed id: 19923724
Date:
11-Jun-09     Release date:   01-Dec-09    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chains
Pfam   ArchSchema ?
P0A6L4  (NANA_ECOLI) -  N-acetylneuraminate lyase
Seq:
Struc:
297 a.a.
298 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 3 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: E.C.4.1.3.3  - N-acetylneuraminate lyase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: N-acetylneuraminate = N-acetyl-D-mannosamine + pyruvate
N-acetylneuraminate
= N-acetyl-D-mannosamine
+
pyruvate
Bound ligand (Het Group name = PYR)
corresponds exactly
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     cytoplasm   1 term 
  Biological process     metabolic process   4 terms 
  Biochemical function     catalytic activity     5 terms  

 

 
    reference    
 
 
Acta Crystallogr Sect F Struct Biol Cryst Commun 65:1088-1090 (2009)
PubMed id: 19923724  
 
 
Structure of an Escherichia coli N-acetyl-D-neuraminic acid lyase mutant, E192N, in complex with pyruvate at 1.45 angstrom resolution.
I.Campeotto, S.B.Carr, C.H.Trinh, A.S.Nelson, A.Berry, S.E.Phillips, A.R.Pearson.
 
  ABSTRACT  
 
The structure of a mutant variant of Escherichia coli N-acetyl-d-neuraminic acid lyase (NAL), E192N, in complex with pyruvate has been determined in a new crystal form. It crystallized in space group P2(1)2(1)2(1), with unit-cell parameters a = 78.3, b = 108.5, c = 148.3 angstrom. Pyruvate has been trapped in the active site as a Schiff base with the catalytic lysine (Lys165) without the need for reduction. Unlike the previously published crystallization conditions for the wild-type enzyme, in which a mother-liquor-derived sulfate ion is strongly bound in the catalytic pocket, the low-salt conditions described here will facilitate the determination of further E. coli NAL structures in complex with other activesite ligands.