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PDBsum entry 3ftn

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protein ligands metals Protein-protein interface(s) links
Oxidoreductase PDB id
3ftn
Jmol
Contents
Protein chains
352 a.a. *
Ligands
ACT ×4
EDO ×11
Metals
_CL ×6
_ZN ×4
Waters ×905
* Residue conservation analysis
PDB id:
3ftn
Name: Oxidoreductase
Title: Q165e/s254k double mutant chimera of alcohol dehydrogenase b of the cofactor binding domain res 153-295 of t. Brockii ad beijerinckii adh
Structure: NADP-dependent alcohol dehydrogenase. Chain: a, b, c, d. Engineered: yes. Mutation: yes. Other_details: residues 153-295 inserted from alcohol dehyd of clostridium beijerinckii
Source: Thermoanaerobacter brockii, clostridiu beijerinckii. Organism_taxid: 29323, 1520. Gene: adh, adh1. Expressed in: escherichia coli. Expression_system_taxid: 562.
Resolution:
2.19Å     R-factor:   0.166     R-free:   0.228
Authors: F.Frolow,E.Goihberg,L.Shimon,Y.Burstein
Key ref: E.Goihberg et al. (2010). Biochemical and structural properties of chimeras constructed by exchange of cofactor-binding domains in alcohol dehydrogenases from thermophilic and mesophilic microorganisms. Biochemistry, 49, 1943-1953. PubMed id: 20102159 DOI: 10.1021/bi901730x
Date:
13-Jan-09     Release date:   26-Jan-10    
PROCHECK
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 Headers
 References

Protein chains
Pfam   ArchSchema ?
P14941  (ADH_THEBR) -  NADP-dependent isopropanol dehydrogenase
Seq:
Struc:
352 a.a.
352 a.a.*
Protein chains
Pfam   ArchSchema ?
P25984  (ADH_CLOBE) -  NADP-dependent isopropanol dehydrogenase
Seq:
Struc:
351 a.a.
352 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 85 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: E.C.1.1.1.80  - Isopropanol dehydrogenase (NADP(+)).
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Propan-2-ol + NADP+ = acetone + NADPH
Propan-2-ol
+ NADP(+)
=
acetone
Bound ligand (Het Group name = ACT)
matches with 40.00% similarity
+ NADPH
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     oxidation-reduction process   1 term 
  Biochemical function     oxidoreductase activity     2 terms  

 

 
    reference    
 
 
DOI no: 10.1021/bi901730x Biochemistry 49:1943-1953 (2010)
PubMed id: 20102159  
 
 
Biochemical and structural properties of chimeras constructed by exchange of cofactor-binding domains in alcohol dehydrogenases from thermophilic and mesophilic microorganisms.
E.Goihberg, M.Peretz, S.Tel-Or, O.Dym, L.Shimon, F.Frolow, Y.Burstein.
 
  ABSTRACT  
 
The cofactor-binding domains (residues 153-295) of the alcohol dehydrogenases from the thermophile Thermoanaerobacter brockii (TbADH), the mesophilic bacterium Clostridium beijerinckii (CbADH), and the protozoan parasite Entamoeba histolytica (EhADH1) have been exchanged. Three chimeras have been constructed. In the first chimera, the cofactor-binding domain of thermophilic TbADH was replaced with the cofactor-binding domain of its mesophilic counterpart CbADH [chimera Chi21((TCT))]. This domain exchange significantly destabilized the parent thermophilic enzyme (DeltaT(1/2) = -18 degrees C). The reverse exchange in CbADH [chimera Chi22((CTC))], however, had little effect on the thermal stability of the parent mesophilic protein. Furthermore, substituting the cofactor-binding domain of TbADH with the homologous domain of EhADH1 [chimera Chi23((TET))] substantially reduced the thermal stability of the thermophilic ADH (DeltaT(1/2) = -51 degrees C) and impeded the oligomerization of the enzyme. All three chimeric proteins and one of their site-directed mutants were crystallized, and their three-dimensional (3D) structures were determined. Comparison of the 3D structures of the chimeras and the chimeric mutant with the structures of their parent ADHs showed no significant changes to their Calpha chains, suggesting that the difference in the thermal stability of the three parent ADHs and their chimeric mutants could be due to a limited number of substitutions located at strategic positions, mainly at the oligomerization interfaces. Indeed, stabilization of the chimeras was achieved, to a significant extent, either by introduction of a proline residue at a strategic position in the major horse liver ADH-type dimerization interface (DeltaT(1/2) = 35 degrees C) or by introduction of intersubunit electrostatic interactions (DeltaT(1/2) = 6 degrees C).