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PDBsum entry 2xsg

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protein ligands metals Protein-protein interface(s) links
Hydrolase PDB id
2xsg
Jmol
Contents
Protein chains
765 a.a.
Ligands
B3P
GOL
Metals
_CA ×2
Waters ×868
PDB id:
2xsg
Name: Hydrolase
Title: Structure of the gh92 family glycosyl hydrolase ccman5
Structure: Ccman5. Chain: a, b. Fragment: residues 1-774. Engineered: yes
Source: Cellulosimicrobium cellulans. Organism_taxid: 1710. Atcc: 27402. Expressed in: escherichia coli. Expression_system_taxid: 469008. Other_details: belgian co-ordinated collections of micro-or strain number lmg 16121
Resolution:
2.00Å     R-factor:   0.193     R-free:   0.238
Authors: E.Baranova,P.Tiels,H.Remaut
Key ref: P.Tiels et al. (2012). A bacterial glycosidase enables mannose-6-phosphate modification and improved cellular uptake of yeast-produced recombinant human lysosomal enzymes. Nat Biotechnol, 30, 1225-1231. PubMed id: 23159880 DOI: 10.1038/nbt.2427
Date:
29-Sep-10     Release date:   12-Oct-11    
PROCHECK
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 Headers
 References

Protein chains
No UniProt id for this chain
Struc:  
Struc: 765 a.a.
Key:    Secondary structure  CATH domain

 Enzyme reactions 
   Enzyme class: E.C.3.2.1.24  - Alpha-mannosidase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Hydrolysis of terminal, non-reducing alpha-D-mannose residues in alpha-D-mannosides.
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     carbohydrate metabolic process   1 term 
  Biochemical function     catalytic activity     2 terms  

 

 
DOI no: 10.1038/nbt.2427 Nat Biotechnol 30:1225-1231 (2012)
PubMed id: 23159880  
 
 
A bacterial glycosidase enables mannose-6-phosphate modification and improved cellular uptake of yeast-produced recombinant human lysosomal enzymes.
P.Tiels, E.Baranova, K.Piens, C.De Visscher, G.Pynaert, W.Nerinckx, J.Stout, F.Fudalej, P.Hulpiau, S.Tännler, S.Geysens, A.Van Hecke, A.Valevska, W.Vervecken, H.Remaut, N.Callewaert.
 
  ABSTRACT  
 
Lysosomal storage diseases are treated with human lysosomal enzymes produced in mammalian cells. Such enzyme therapeutics contain relatively low levels of mannose-6-phosphate, which is required to target them to the lysosomes of patient cells. Here we describe a method for increasing mannose-6-phosphate modification of lysosomal enzymes produced in yeast. We identified a glycosidase from C. cellulans that 'uncaps' N-glycans modified by yeast-type mannose-Pi-6-mannose to generate mammalian-type N-glycans with a mannose-6-phosphate substitution. Determination of the crystal structure of this glycosidase provided insight into its substrate specificity. We used this uncapping enzyme together with α-mannosidase to produce in yeast a form of the Pompe disease enzyme α-glucosidase rich in mannose-6-phosphate. Compared with the currently used therapeutic version, this form of α-glucosidase was more efficiently taken up by fibroblasts from Pompe disease patients, and it more effectively reduced cardiac muscular glycogen storage in a mouse model of the disease.