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PDBsum entry 3m0v

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protein ligands metals Protein-protein interface(s) links
Isomerase PDB id
3m0v
Jmol
Contents
Protein chains
421 a.a. *
Ligands
RNS ×4
Metals
_MN ×8
Waters ×1745
* Residue conservation analysis
PDB id:
3m0v
Name: Isomerase
Title: Crystal structure of pseudomonas stutzeri l-rhamnose isomera s329l in complex with l-rhamnose
Structure: L-rhamnose isomerase. Chain: a, b, c, d. Engineered: yes. Mutation: yes
Source: Pseudomonas stutzeri. Pseudomonas perfectomarina. Organism_taxid: 316. Expressed in: escherichia coli. Expression_system_taxid: 562.
Resolution:
1.79Å     R-factor:   0.183     R-free:   0.216
Authors: H.Yoshida,K.Takeda,K.Izumori,S.Kamitori
Key ref: H.Yoshida et al. (2010). Elucidation of the role of Ser329 and the C-terminal region in the catalytic activity of Pseudomonas stutzeri L-rhamnose isomerase. Protein Eng Des Sel, 23, 919-927. PubMed id: 20977999
Date:
03-Mar-10     Release date:   10-Nov-10    
PROCHECK
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 Headers
 References

Protein chains
Pfam   ArchSchema ?
Q75WH8  (Q75WH8_PSEST) -  L-rhamnose isomerase
Seq:
Struc:
430 a.a.
421 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 2 residue positions (black crosses)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     metabolic process   1 term 
  Biochemical function     isomerase activity     2 terms  

 

 
Protein Eng Des Sel 23:919-927 (2010)
PubMed id: 20977999  
 
 
Elucidation of the role of Ser329 and the C-terminal region in the catalytic activity of Pseudomonas stutzeri L-rhamnose isomerase.
H.Yoshida, K.Takeda, K.Izumori, S.Kamitori.
 
  ABSTRACT  
 
Pseudomonas stutzeri l-rhamnose isomerase (l-RhI) is capable of catalyzing the isomerization between various aldoses and ketoses, showing high catalytic activity with broad substrate-specificity compared with Escherichia coli l-RhI. In a previous study, the crystal structure of P. stutzeri l-RhI revealed an active site comparable with that of E. coli l-RhI and d-xylose isomerases (d-XIs) with structurally conserved amino acids, but also with a different residue seemingly responsible for the specificity of P. stutzeri l-RhI, though the residue itself does not interact with the bound substrate. This residue, Ser329, corresponds to Phe336 in E. coli l-RhI and Lys294 in Actinoplanes missouriensis d-XI. To elucidate the role of Ser329 in P. stutzeri l-RhI, we constructed mutants, S329F (E. coli l-RhI type), S329K (A. missouriensis d-XI type), S329L and S329A. Analyses of the catalytic activity and crystal structure of the mutants revealed a hydroxyl group of Ser329 to be crucial for catalytic activity via interaction with a water molecule. In addition, in complexes with substrate, the mutants S329F and S329L exhibited significant electron density in the C-terminal region not observed in the wild-type P. stutzeri l-RhI. The C-terminal region of P. stutzeri l-RhI has flexibility and shows a flip-flop movement at the inter-molecular surface of the dimeric form.