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PDBsum entry 1egz

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protein metals Protein-protein interface(s) links
Hydrolase PDB id
1egz
Jmol
Contents
Protein chains
291 a.a. *
Metals
_CA ×3
Waters ×333
* Residue conservation analysis
PDB id:
1egz
Name: Hydrolase
Title: Cellulase cel5 from erwinia chrysanthemi, a family gh 5-2 enzyme
Structure: Endoglucanase z. Chain: a, b, c. Fragment: catalytic domain. Synonym: egz, cel5. Other_details: three copies in the asymmetric unit
Source: Erwinia chrysanthemi. Organism_taxid: 556. Secretion: type ii. Cellular_location: exterior. Other_details: gene: celz
Resolution:
2.30Å     R-factor:   0.179     R-free:   0.263
Authors: M.Czjzek,M.El Hassouni,B.Py,F.Barras
Key ref:
V.Chapon et al. (2001). Type II protein secretion in gram-negative pathogenic bacteria: the study of the structure/secretion relationships of the cellulase Cel5 (formerly EGZ) from Erwinia chrysanthemi. J Mol Biol, 310, 1055-1066. PubMed id: 11501995 DOI: 10.1006/jmbi.2001.4787
Date:
18-Mar-99     Release date:   26-Mar-99    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chains
Pfam   ArchSchema ?
P07103  (GUNZ_DICD3) -  Endoglucanase Z
Seq:
Struc:
426 a.a.
291 a.a.
Key:    PfamA domain  Secondary structure  CATH domain

 Enzyme reactions 
   Enzyme class: E.C.3.2.1.4  - Cellulase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Endohydrolysis of 1,4-beta-D-glucosidic linkages in cellulose, lichenin and cereal beta-D-glucans.
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     carbohydrate metabolic process   1 term 
  Biochemical function     hydrolase activity, hydrolyzing O-glycosyl compounds     1 term  

 

 
DOI no: 10.1006/jmbi.2001.4787 J Mol Biol 310:1055-1066 (2001)
PubMed id: 11501995  
 
 
Type II protein secretion in gram-negative pathogenic bacteria: the study of the structure/secretion relationships of the cellulase Cel5 (formerly EGZ) from Erwinia chrysanthemi.
V.Chapon, M.Czjzek, M.El Hassouni, B.Py, M.Juy, F.Barras.
 
  ABSTRACT  
 
Erwinia chrysanthemi, a Gram-negative plant pathogen, secretes the cellulase Cel5 (formerly EGZ) via the type II secretion pathway (referred to as Out). Cel5 is composed of two domains, a large N-terminal catalytic domain (390 amino acid residues) and a small C-terminal cellulose-binding domain (62 amino acid residues) separated by a linker region. A combination of mutagenesis and structural analysis permitted us to investigate the structure/secretion relationships with respect to the catalytic domain of Cel5. The 3D structure of the catalytic domain was solved by molecular replacement at 2.3 A resolution. Cel5 exhibits the (beta/alpha)8 structural fold and two extra-barrel features. Our previous genetic study based upon tRNA-mediated suppression allowed us to predict positions of importance in the molecule in relation to structure and catalysis. Remarkably, all of the predictions proved to be correct when compared with the present structural information. Mutations of Arg57, which is located at the heart of the catalytic domain, allowed us to test the consequences of structural modifications on the secretion efficiency. The results revealed that secretability imposes remarkably strong constraints upon folding. In particular, an Arg-to-His mutation yielded a species that folded to a stable conformation close to, but distinct from the wild-type, which however was not secretable. We discuss the relationships between folding of a protein in the periplasm, en route to the cell exterior, and presentation of secretion information. We propose that different solutions have been selected for type II secreted exoproteins in order to meet the constraints imposed by their interaction with their respective secretion machineries. We propose that evolutionary pressure has led to the adaptation of different secretion motifs for different type II exoproteins.
 
  Selected figure(s)  
 
Figure 2.
Figure 2. Close-up view of the catalytic site and the highly conserved residues of Cel5 from E. chrysanthemi (yellow), and of Cel5A from B. agharadherans (magenta). The numbers correspond to the residues of Cel5 from E. chrysanthemi. (a) His100 is replaced by a leucine residue in Cel5A from B. agaradherans. (b) The NH group of Trp28 in Cel5 superposes well with the hydroxyl group of Tyr66, although they are donated by different stretches of the b-barrel. (c) His198 is turned towards the active site in Cel5 from E. chrysanthemi, whereas His206 points into the solvent in the structural model of Cel5A of B. agaradherans. The Figures were produced with TURBO-FRODO[45].
Figure 3.
Figure 3. (a) Ribbon representation of the (b/a)[8] barrel highlighting the positions of all ten arginine residues (balls-and-sticks representation) of Cel5. (b) A close-up view of Arg57 in the heart of the catalytic domain, displaying the hydrogen bonding network in which Arg57 is involved. Connected residues are His98, Glu129, Asn132 and Glu220, which are all related directly to catalysis, and Ser25 and Thr219, which participate in structural stabilization. The Figures were produced using the programs MOLSCRIPT [51] and Raster3D [52].
 
  The above figures are reprinted by permission from Elsevier: J Mol Biol (2001, 310, 1055-1066) copyright 2001.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
20525826 A.Filloux (2010).
Secretion signal and protein targeting in bacteria: a biological puzzle.
  J Bacteriol, 192, 3847-3849.  
19817866 M.M.Pearce, and N.P.Cianciotto (2009).
Legionella pneumophila secretes an endoglucanase that belongs to the family-5 of glycosyl hydrolases and is dependent upon type II secretion.
  FEMS Microbiol Lett, 300, 256-264.  
19304855 S.J.Krebs, T.J.Kirn, and R.K.Taylor (2009).
Genetic mapping of secretion and functional determinants of the Vibrio cholerae TcpF colonization factor.
  J Bacteriol, 191, 3665-3676.  
18978048 V.Douet, D.Expert, F.Barras, and B.Py (2009).
Erwinia chrysanthemi iron metabolism: the unexpected implication of the inner membrane platform within the type II secretion system.
  J Bacteriol, 191, 795-804.  
18218035 F.F.Evans, S.Egan, and S.Kjelleberg (2008).
Ecology of type II secretion in marine gammaproteobacteria.
  Environ Microbiol, 10, 1101-1107.  
17209016 A.Lykidis, K.Mavromatis, N.Ivanova, I.Anderson, M.Land, G.DiBartolo, M.Martinez, A.Lapidus, S.Lucas, A.Copeland, P.Richardson, D.B.Wilson, and N.Kyrpides (2007).
Genome sequence and analysis of the soil cellulolytic actinomycete Thermobifida fusca YX.
  J Bacteriol, 189, 2477-2486.  
16630257 A.M.Cusano, E.Parrilli, A.Duilio, G.Sannia, G.Marino, and M.L.Tutino (2006).
Secretion of psychrophilic alpha-amylase deletion mutants in Pseudoalteromonas haloplanktis TAC125.
  FEMS Microbiol Lett, 258, 67-71.  
16240096 E.Papaleo, P.Fantucci, M.Vai, and L.De Gioia (2006).
Three-dimensional structure of the catalytic domain of the yeast beta-(1,3)-glucan transferase Gas1: a molecular modeling investigation.
  J Mol Model, 12, 237-248.  
16704357 I.K.Toth, L.Pritchard, and P.R.Birch (2006).
Comparative genomics reveals what makes an enterobacterial plant pathogen.
  Annu Rev Phytopathol, 44, 305-336.  
15808747 G.M.Preston, D.J.Studholme, and I.Caldelari (2005).
Profiling the secretomes of plant pathogenic Proteobacteria.
  FEMS Microbiol Rev, 29, 331-360.  
16216510 N.P.Cianciotto (2005).
Type II secretion: a protein secretion system for all seasons.
  Trends Microbiol, 13, 581-588.  
16199575 O.Francetić, and A.P.Pugsley (2005).
Towards the identification of type II secretion signals in a nonacylated variant of pullulanase from Klebsiella oxytoca.
  J Bacteriol, 187, 7045-7055.  
15666145 W.J.Lim, S.Y.Hong, C.L.An, K.M.Cho, B.R.Choi, Y.K.Kim, J.M.An, J.M.Kang, S.M.Lee, S.J.Cho, H.Kim, and H.D.Yun (2005).
Construction of minimum size cellulase (Cel5Z) from Pectobacterium chrysanthemi PY35 by removal of the C-terminal region.
  Appl Microbiol Biotechnol, 68, 46-52.  
15604820 L.Hildén, and G.Johansson (2004).
Recent developments on cellulases and carbohydrate-binding modules with cellulose affinity.
  Biotechnol Lett, 26, 1683-1693.  
12832777 S.Violot, R.Haser, G.Sonan, D.Georlette, G.Feller, and N.Aghajari (2003).
Expression, purification, crystallization and preliminary X-ray crystallographic studies of a psychrophilic cellulase from Pseudoalteromonas haloplanktis.
  Acta Crystallogr D Biol Crystallogr, 59, 1256-1258.  
12067351 M.Gérard-Vincent, V.Robert, G.Ball, S.Bleves, G.P.Michel, A.Lazdunski, and A.Filloux (2002).
Identification of XcpP domains that confer functionality and specificity to the Pseudomonas aeruginosa type II secretion apparatus.
  Mol Microbiol, 44, 1651-1665.  
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.