PDBsum entry 1c8x

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Hydrolase PDB id
Protein chain
265 a.a. *
Waters ×152
* Residue conservation analysis
PDB id:
Name: Hydrolase
Title: Endo-beta-n-acetylglucosaminidase h, d130e mutant
Structure: Endo-beta-n-acetylglucosaminidase h. Chain: a. Engineered: yes. Mutation: yes
Source: Streptomyces plicatus. Organism_taxid: 1922. Expressed in: escherichia coli. Expression_system_taxid: 562.
2.00Å     R-factor:   0.182    
Authors: V.Rao,C.Tao,C.Guan,P.Van Roey
Key ref: V.Rao et al. (1999). Mutations of endo-beta-N-acetylglucosaminidase H active site residueAs sp130 anG glu132: activities and conformations. Protein Sci, 8, 2338-2346. PubMed id: 10595536 DOI: 10.1110/ps.8.11.2338
30-Jul-99     Release date:   26-Nov-99    
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Protein chain
Pfam   ArchSchema ?
P04067  (EBAG_STRPL) -  Endo-beta-N-acetylglucosaminidase H
313 a.a.
265 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Enzyme reactions 
   Enzyme class: E.C.  - Mannosyl-glycoprotein endo-beta-N-acetylglucosaminidase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Endohydrolysis of the di-N-acetylchitobiosyl unit in high-mannose glycopeptides and glycoproteins containing the -[Man(GlcNAc)2]Asn- structure. One N-acetyl-D-glucosamine residue remains attached to the protein; the rest of the oligosaccharide is released intact.
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     metabolic process   2 terms 
  Biochemical function     hydrolase activity     4 terms  


DOI no: 10.1110/ps.8.11.2338 Protein Sci 8:2338-2346 (1999)
PubMed id: 10595536  
Mutations of endo-beta-N-acetylglucosaminidase H active site residueAs sp130 anG glu132: activities and conformations.
V.Rao, T.Cui, C.Guan, P.Van Roey.
Endo-beta-N-acetylglucosaminidase H hydrolyzes the beta-(1-4)-glycosidic link of the N,N'-diacetylchitobiose core of high-mannose and hybrid asparagine-linked oligosaccharides. Seven mutants of the active site residues, Asp130 and Glu132, have been prepared, assayed, and crystallized. They include single site mutants of each residue to the corresponding amide, to Ala and to the alternate acidic residue, and to the double amide mutant. The mutants of Asp130 are more active than the corresponding Glu132 mutants, consistent with the assignment of the latter residue as the primary catalytic residue. The amide mutants are more active than the alternate acidic residue mutants, which in turn are more active than the Ala mutants. The structures of the Asn mutant of Asp130 and the double mutant are very similar to that of the wild-type enzyme. Several residues surrounding the mutated residues, including some that form part of the core of the beta-barrel and especially Tyr168 and Tyr244, adopt a very different conformation in the structures of the other two mutants of Asp130 and in the Asp mutant of Glu132. The results show that the residues in the upper layers of the beta-barrel can organize into two very distinct packing arrangements that depend on subtle electrostatic and steric differences and that greatly affect the geometry of the substrate-binding cleft. Consequently, the relative activities of several of the mutants are defined by structural changes, leading to impaired substrate binding, in addition to changes in functionality.

Literature references that cite this PDB file's key reference

  PubMed id Reference
20084296 H.Li, and L.H.Greene (2010).
Sequence and structural analysis of the chitinase insertion domain reveals two conserved motifs involved in chitin-binding.
  PLoS One, 5, e8654.  
19201751 L.Zhang, A.G.Bharadwaj, A.Casper, J.Barkley, J.J.Barycki, and M.A.Simpson (2009).
Hyaluronidase activity of human Hyal1 requires active site acidic and tyrosine residues.
  J Biol Chem, 284, 9433-9442.  
17543889 Zaheer-ul-Haq, P.Dalal, N.N.Aronson, and J.D.Madura (2007).
Family 18 chitolectins: comparison of MGP40 and HUMGP39.
  Biochem Biophys Res Commun, 359, 221-226.  
12114544 T.Suzuki, K.Yano, S.Sugimoto, K.Kitajima, W.J.Lennarz, S.Inoue, Y.Inoue, and Y.Emori (2002).
Endo-beta-N-acetylglucosaminidase, an enzyme involved in processing of free oligosaccharides in the cytosol.
  Proc Natl Acad Sci U S A, 99, 9691-9696.  
11515537 K.Fujita, R.Nakatake, K.Yamabe, A.Watanabe, Y.Asada, and K.Takegawa (2001).
Identification of amino acid residues essential for the substrate specificity of Flavobacterium sp. endo-beta-N-acetylglucosaminidase.
  Biosci Biotechnol Biochem, 65, 1542-1548.  
10891067 C.A.Waddling, T.H.Plummer, A.L.Tarentino, and P.Van Roey (2000).
Structural basis for the substrate specificity of endo-beta-N-acetylglucosaminidase F(3).
  Biochemistry, 39, 7878-7885.
PDB codes: 1eok 1eom
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