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PDBsum entry 1mw0

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protein ligands links
Transferase PDB id
1mw0
Jmol
Contents
Protein chain
628 a.a. *
Ligands
GLC-GLC-GLC-GLC-
GLC-GLC-GLC
×2
BGC-GLC-GLC-GLC
DTT
Waters ×411
* Residue conservation analysis
PDB id:
1mw0
Name: Transferase
Title: Amylosucrase mutant e328q co-crystallized with maltoheptaose then soaked with maltoheptaose.
Structure: Amylosucrase. Chain: a. Engineered: yes. Mutation: yes
Source: Neisseria polysaccharea. Organism_taxid: 489. Expressed in: escherichia coli. Expression_system_taxid: 562.
Resolution:
2.01Å     R-factor:   0.185     R-free:   0.223
Authors: L.K.Skov,O.Mirza,D.Sprogoe,I.Dar,M.Remaud-Simeon,C.Albenne, P.Monsan,M.Gajhede
Key ref:
L.K.Skov et al. (2002). Oligosaccharide and sucrose complexes of amylosucrase. Structural implications for the polymerase activity. J Biol Chem, 277, 47741-47747. PubMed id: 12364331 DOI: 10.1074/jbc.M207860200
Date:
27-Sep-02     Release date:   18-Dec-02    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam   ArchSchema ?
Q9ZEU2  (AMYS_NEIPO) -  Amylosucrase
Seq:
Struc:
 
Seq:
Struc:
636 a.a.
628 a.a.*
Key:    PfamA domain  PfamB domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 6 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: E.C.2.4.1.4  - Amylosucrase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Sucrose + ((1->4)-alpha-D-glucosyl)(n) = D-fructose + ((1->4)-alpha-D- glucosyl)(n+1)
Sucrose
+ ((1->4)-alpha-D-glucosyl)(n)
=
D-fructose
Bound ligand (Het Group name = GLC)
corresponds exactly
+ ((1->4)-alpha-D- glucosyl)(n+1)
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     extracellular region   1 term 
  Biological process     carbohydrate metabolic process   1 term 
  Biochemical function     catalytic activity     5 terms  

 

 
    Added reference    
 
 
DOI no: 10.1074/jbc.M207860200 J Biol Chem 277:47741-47747 (2002)
PubMed id: 12364331  
 
 
Oligosaccharide and sucrose complexes of amylosucrase. Structural implications for the polymerase activity.
L.K.Skov, O.Mirza, D.Sprogøe, I.Dar, M.Remaud-Simeon, C.Albenne, P.Monsan, M.Gajhede.
 
  ABSTRACT  
 
The glucosyltransferase amylosucrase is structurally quite similar to the hydrolase alpha-amylase. How this switch in functionality is achieved is an important and fundamental question. The inactive E328Q amylosucrase variant has been co-crystallized with maltoheptaose, and the structure was determined by x-ray crystallography to 2.2 A resolution, revealing a maltoheptaose binding site in the B'-domain somewhat distant from the active site. Additional soaking of these crystals with maltoheptaose resulted in replacement of Tris in the active site with maltoheptaose, allowing the mapping of the -1 to +5 binding subsites. Crystals of amylosucrase were soaked with sucrose at different concentrations. The structures at approximately 2.1 A resolution revealed three new binding sites of different affinity. The highest affinity binding site is close to the active site but is not in the previously identified substrate access channel. Allosteric regulation seems necessary to facilitate access from this binding site. The structures show the pivotal role of the B'-domain in the transferase reaction. Based on these observations, an extension of the hydrolase reaction mechanism valid for this enzyme can be proposed. In this mechanism, the glycogen-like polymer is bound in the widest access channel to the active site. The polymer binding introduces structural changes that allow sucrose to migrate from its binding site into the active site and displace the polymer.
 
  Selected figure(s)  
 
Figure 1.
Fig. 1. Stereo picture illustrating the various sucrose- and oligosaccharide binding sites on amylosucrase. The three oligosacchararide binding sites are labeled OB1, OB2, and OB3. Sucrose at the surface site is colored magenta and labeled SB2. The B'-domain is colored yellow.
Figure 5.
Fig. 5. Schematic representation of the interactions at the AS·sucrose binding site (SB2).
 
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (2002, 277, 47741-47747) copyright 2002.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
21113589 J.Schneider, C.Fricke, H.Overwin, and B.Hofer (2011).
High level expression of a recombinant amylosucrase gene and selected properties of the enzyme.
  Appl Microbiol Biotechnol, 89, 1821-1829.  
19966417 E.Champion, M.Remaud-Simeon, L.K.Skov, J.S.Kastrup, M.Gajhede, and O.Mirza (2009).
The apo structure of sucrose hydrolase from Xanthomonas campestris pv. campestris shows an open active-site groove.
  Acta Crystallogr D Biol Crystallogr, 65, 1309-1314.
PDB code: 2wpg
19801480 J.Schneider, C.Fricke, H.Overwin, B.Hofmann, and B.Hofer (2009).
Generation of amylosucrase variants that terminate catalysis of acceptor elongation at the di- or trisaccharide stage.
  Appl Environ Microbiol, 75, 7453-7460.  
18522649 S.Emond, S.Mondeil, K.Jaziri, I.André, P.Monsan, M.Remaud-Siméon, and G.Potocki-Véronèse (2008).
Cloning, purification and characterization of a thermostable amylosucrase from Deinococcus geothermalis.
  FEMS Microbiol Lett, 285, 25-32.  
17044042 C.Albenne, L.K.Skov, V.Tran, M.Gajhede, P.Monsan, M.Remaud-Siméon, and G.André-Leroux (2007).
Towards the molecular understanding of glycogen elongation by amylosucrase.
  Proteins, 66, 118-126.  
17803687 S.Bozonnet, M.T.Jensen, M.M.Nielsen, N.Aghajari, M.H.Jensen, B.Kramhøft, M.Willemoës, S.Tranier, R.Haser, and B.Svensson (2007).
The 'pair of sugar tongs' site on the non-catalytic domain C of barley alpha-amylase participates in substrate binding and activity.
  FEBS J, 274, 5055-5067.
PDB codes: 2qps 2qpu
16441655 B.A.van der Veen, L.K.Skov, G.Potocki-Véronèse, M.Gajhede, P.Monsan, and M.Remaud-Simeon (2006).
Increased amylosucrase activity and specificity, and identification of regions important for activity, specificity and stability through molecular evolution.
  FEBS J, 273, 673-681.  
16524921 S.A.van Hijum, S.Kralj, L.K.Ozimek, L.Dijkhuizen, and I.G.van Geel-Schutten (2006).
Structure-function relationships of glucansucrase and fructansucrase enzymes from lactic acid bacteria.
  Microbiol Mol Biol Rev, 70, 157-176.  
15062085 J.Allouch, W.Helbert, B.Henrissat, and M.Czjzek (2004).
Parallel substrate binding sites in a beta-agarase suggest a novel mode of action on double-helical agarose.
  Structure, 12, 623-632.
PDB code: 1urx
15027107 J.Cortés, T.Siméon, M.Remaud-Siméon, and V.Tran (2004).
Geometric algorithms for the conformational analysis of long protein loops.
  J Comput Chem, 25, 956-967.  
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 code is shown on the right.