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PDBsum entry 2wil

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protein ligands metals Protein-protein interface(s) links
Hydrolase PDB id
2wil
Jmol
Contents
Protein chain
526 a.a. *
Ligands
TCX ×2
NAG-NAG-FUC ×4
NAG ×6
SO4 ×3
Metals
_CL
Waters ×137
* Residue conservation analysis
PDB id:
2wil
Name: Hydrolase
Title: Aged form of human butyrylcholinesterase inhibited by tabun analogue ta5
Structure: Cholinesterase. Chain: a, b. Fragment: residues 29-557. Synonym: acylcholine acylhydrolase, butyrylcholine esterase, choline esterase ii, pseudocholinesterase. Engineered: yes. Mutation: yes. Other_details: s198 is phosphoramidylated
Source: Homo sapiens. Human. Organism_taxid: 9606. Expressed in: cricetulus griseus. Expression_system_taxid: 10029. Expression_system_cell_line: ovary cells.
Resolution:
3.10Å     R-factor:   0.221     R-free:   0.252
Authors: E.Carletti,N.Aurbek,E.Gillon,M.Loiodice,Y.Nicolet, J.Fontecilla,P.Masson,H.Thiermann,F.Nachon,F.Worek
Key ref: E.Carletti et al. (2009). Structure-activity analysis of aging and reactivation of human butyrylcholinesterase inhibited by analogues of tabun. Biochem J, 421, 97. PubMed id: 19368529
Date:
12-May-09     Release date:   26-May-09    
PROCHECK
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 Headers
 References

Protein chains
Pfam   ArchSchema ?
P06276  (CHLE_HUMAN) -  Cholinesterase
Seq:
Struc:
 
Seq:
Struc:
602 a.a.
526 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 2 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: E.C.3.1.1.8  - Cholinesterase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: An acylcholine + H2O = choline + a carboxylate
acylcholine
+ H(2)O
= choline
+ carboxylate
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     extracellular region   7 terms 
  Biological process     response to drug   13 terms 
  Biochemical function     catalytic activity     9 terms  

 

 
    reference    
 
 
Biochem J 421:97 (2009)
PubMed id: 19368529  
 
 
Structure-activity analysis of aging and reactivation of human butyrylcholinesterase inhibited by analogues of tabun.
E.Carletti, N.Aurbek, E.Gillon, M.Loiodice, Y.Nicolet, J.C.Fontecilla-Camps, P.Masson, H.Thiermann, F.Nachon, F.Worek.
 
  ABSTRACT  
 
hBChE [human BChE (butyrylcholinesterase)] naturally scavenges OPs (organophosphates). This bioscavenger is currently in Clinical Phase I for pretreatment of OP intoxication. Phosphylated ChEs (cholinesterases) can undergo a spontaneous time-dependent process called 'aging' during which the conjugate is dealkylated, leading to creation of an enzyme that cannot be reactivated. hBChE inhibited by phosphoramidates such as tabun displays a peculiar resistance to oxime-mediated reactivation. We investigated the basis of oxime resistance of phosphoramidyl-BChE conjugates by determining the kinetics of inhibition, reactivation (obidoxime {1,1'-(oxybis-methylene) bis[4-(hydroxyimino) methyl] pyridinium dichloride}, TMB-4 [1,3-trimethylene-bis(4-hydroxyiminomethylpyridinium) dibromide], HLö 7 {1-[[[4-(aminocarbonyl) pyridinio]methoxy]methyl]-2,4-bis-[(hydroxyimino)methyl] pyridinium dimethanesulfonate)}, HI-6 {1-[[[4-(aminocarbonyl) pyridinio] methoxy] methyl]-2-[(hydroxyimino)methyl]pyridinium dichloride monohydrate} and aging, and the crystal structures of hBChE inhibited by different N-monoalkyl and N,N-dialkyl tabun analogues. The refined structures of aged hBChE conjugates show that aging proceeds through O-dealkylation of the P(R) enantiomer of N,N-diethyl and N-propyl analogues, with subsequent formation of a salt bridge preventing reactivation, similarly to a previous observation made on tabun-ChE conjugates. Interestingly, the N-methyl analogue projects its amino group towards the choline-binding pocket, so that aging proceeds through deamination. This orientation results from a preference of hBChE's acyl-binding pocket for larger than 2-atoms linear substituents. The correlation between the inhibitory potency and the N-monoalkyl chain length is related to increasingly optimized interactions with the acyl-binding pocket as shown by the X-ray structures. These kinetics and X-ray data lead to a structure-activity relationship that highlights steric and electronic effects of the amino substituent of phosphoramidate. This study provides the structural basis to design new oximes capable of reactivating phosphoramidyl-hBChE conjugates after intoxication, notably when hBChE is used as pretreatment, or to design BChE-based catalytic bioscavengers.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
21091433 F.Nachon, E.Carletti, M.Wandhammer, Y.Nicolet, L.M.Schopfer, P.Masson, and O.Lockridge (2011).
X-ray crystallographic snapshots of reaction intermediates in the G117H mutant of human butyrylcholinesterase, a nerve agent target engineered into a catalytic bioscavenger.
  Biochem J, 434, 73-82.
PDB codes: 2xmb 2xmc 2xmd 2xmg
21504805 L.Wang, D.Du, D.Lu, C.T.Lin, J.N.Smith, C.Timchalk, F.Liu, J.Wang, and Y.Lin (2011).
Enzyme-linked immunosorbent assay for detection of organophosphorylated butyrylcholinesterase: a biomarker of exposure to organophosphate agents.
  Anal Chim Acta, 693, 1-6.  
20004171 P.Masson, and O.Lockridge (2010).
Butyrylcholinesterase for protection from organophosphorus poisons: catalytic complexities and hysteretic behavior.
  Arch Biochem Biophys, 494, 107-120.  
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