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Transferase PDB id
2cqs
Jmol
Contents
Protein chains
822 a.a. *
Ligands
BGC ×2
SO4 ×2
Waters ×1074
* Residue conservation analysis
PDB id:
2cqs
Name: Transferase
Title: Crystal structure of cellvibrio gilvus cellobiose phosphorylase crystallized from ammonium sulfate
Structure: Cellobiose phosphorylase. Chain: a, b. Engineered: yes
Source: Cellvibrio gilvus. Organism_taxid: 11. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.
Biol. unit: Dimer (from PQS)
Resolution:
2.00Å     R-factor:   0.176     R-free:   0.213
Authors: M.Hidaka,M.Kitaoka,K.Hayashi,T.Wakagi,H.Shoun,S.Fushinobu
Key ref: M.Hidaka et al. (2006). Structural dissection of the reaction mechanism of cellobiose phosphorylase. Biochem J, 398, 37-43. PubMed id: 16646954 DOI: 10.1042/BJ20060274
Date:
20-May-05     Release date:   16-May-06    
PROCHECK
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 Headers
 References

Protein chains
Pfam   ArchSchema ?
O66264  (O66264_9GAMM) -  Cellobiose Phosphorylase
Seq:
Struc: 		 
Seq:
Struc: 		822 a.a.
822 a.a.
Key:    PfamA domain  Secondary structure

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

 

 
DOI no: 10.1042/BJ20060274 Biochem J 398:37-43 (2006)
PubMed id: 16646954  
 
 
Structural dissection of the reaction mechanism of cellobiose phosphorylase.
M.Hidaka, M.Kitaoka, K.Hayashi, T.Wakagi, H.Shoun, S.Fushinobu.
 
  ABSTRACT  
 
Cellobiose phosphorylase, a member of the glycoside hydrolase family 94, catalyses the reversible phosphorolysis of cellobiose into alpha-D-glucose 1-phosphate and D-glucose with inversion of the anomeric configuration. The substrate specificity and reaction mechanism of cellobiose phosphorylase from Cellvibrio gilvus have been investigated in detail. We have determined the crystal structure of the glucose-sulphate and glucose-phosphate complexes of this enzyme at a maximal resolution of 2.0 A (1 A=0.1 nm). The phosphate ion is strongly held through several hydrogen bonds, and the configuration appears to be suitable for direct nucleophilic attack to an anomeric centre. Structural features around the sugar-donor and sugar-acceptor sites were consistent with the results of extensive kinetic studies. When we compared this structure with that of homologous chitobiose phosphorylase, we identified key residues for substrate discrimination between glucose and N-acetylglucosamine in both the sugar-donor and sugar-acceptor sites. We found that the active site pocket of cellobiose phosphorylase was covered by an additional loop, indicating that some conformational change is required upon substrate binding. Information on the three-dimensional structure of cellobiose phosphorylase will facilitate engineering of this enzyme, the application of which to practical oligosaccharide synthesis has already been established.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
21344678 G.Hai Tran, T.Desmet, M.R.De Groeve, and W.Soetaert (2011).
Probing the active site of cellodextrin phosphorylase from Clostridium stercorarium: Kinetic characterization, ligand docking, and site-directed mutagenesis.
  Biotechnol Prog, 27, 326-332.  
21154671 C.Luley-Goedl, and B.Nidetzky (2010).
Carbohydrate synthesis by disaccharide phosphorylases: reactions, catalytic mechanisms and application in the glycosciences.
  Biotechnol J, 5, 1324-1338.  
20713411 H.Nakai, B.O.Petersen, Y.Westphal, A.Dilokpimol, M.Abou Hachem, J.Ã.˜.Duus, H.A.Schols, and B.Svensson (2010).
Rational engineering of Lactobacillus acidophilus NCFM maltose phosphorylase into either trehalose or kojibiose dual specificity phosphorylase.
  Protein Eng Des Sel, 23, 781-787.  
20517986 M.R.De Groeve, L.Remmery, A.Van Hoorebeke, J.Stout, T.Desmet, S.N.Savvides, and W.Soetaert (2010).
Construction of cellobiose phosphorylase variants with broadened acceptor specificity towards anomerically substituted glucosides.
  Biotechnol Bioeng, 107, 413-420.  
21150123 S.Fushinobu (2010).
Unique sugar metabolic pathways of bifidobacteria.
  Biosci Biotechnol Biochem, 74, 2374-2384.  
20552664 T.V.Vuong, and D.B.Wilson (2010).
Glycoside hydrolases: catalytic base/nucleophile diversity.
  Biotechnol Bioeng, 107, 195-205.  
19124470 M.Hidaka, M.Nishimoto, M.Kitaoka, T.Wakagi, H.Shoun, and S.Fushinobu (2009).
The Crystal Structure of Galacto-N-biose/Lacto-N-biose I Phosphorylase: A LARGE DEFORMATION OF A TIM BARREL SCAFFOLD.
  J Biol Chem, 284, 7273-7283.
PDB codes: 2zus 2zut 2zuu 2zuv 2zuw
19491100 M.Nakajima, M.Nishimoto, and M.Kitaoka (2009).
Characterization of Three {beta}-Galactoside Phosphorylases from Clostridium phytofermentans: DISCOVERY OF D-GALACTOSYL-{beta}1->4-L-RHAMNOSE PHOSPHORYLASE.
  J Biol Chem, 284, 19220-19227.  
19487233 M.R.De Groeve, M.De Baere, L.Hoflack, T.Desmet, E.J.Vandamme, and W.Soetaert (2009).
Creating lactose phosphorylase enzymes by directed evolution of cellobiose phosphorylase.
  Protein Eng Des Sel, 22, 393-399.  
17587697 M.Nishimoto, and M.Kitaoka (2007).
Identification of the putative proton donor residue of lacto-N-biose phosphorylase (EC 2.4.1.211).
  Biosci Biotechnol Biochem, 71, 1587-1591.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB codes are shown on the right.