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PDBsum entry 2e0w

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protein Protein-protein interface(s) links
Transferase PDB id
2e0w
Jmol
Contents
Protein chains
496 a.a. *
Waters ×53
* Residue conservation analysis
PDB id:
2e0w
Name: Transferase
Title: T391a precursor mutant protein of gamma-glutamyltranspeptida escherichia coli
Structure: Gamma-glutamyltranspeptidase. Chain: a, b. Synonym: gamma-glutamyltransferase large subunit and small engineered: yes. Mutation: yes
Source: Escherichia coli k12. Organism_taxid: 83333. Strain: k-12. Gene: ggt. Expressed in: escherichia coli. Expression_system_taxid: 562.
Biol. unit: Dimer (from PQS)
Resolution:
2.55Å     R-factor:   0.220     R-free:   0.270
Authors: T.Okada,K.Wada,K.Fukuyama
Key ref:
T.Okada et al. (2007). Crystal structure of the gamma-glutamyltranspeptidase precursor protein from Escherichia coli. Structural changes upon autocatalytic processing and implications for the maturation mechanism. J Biol Chem, 282, 2433-2439. PubMed id: 17135273 DOI: 10.1074/jbc.M607490200
Date:
16-Oct-06     Release date:   28-Nov-06    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chains
Pfam   ArchSchema ?
P18956  (GGT_ECOLI) -  Gamma-glutamyltranspeptidase
Seq:
Struc:
 
Seq:
Struc:
580 a.a.
496 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Enzyme reactions 
   Enzyme class 2: E.C.2.3.2.2  - Gamma-glutamyltransferase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: A (5-L-glutamyl)-peptide + an amino acid = a peptide + a 5-L-glutamyl amino acid
(5-L-glutamyl)-peptide
+ amino acid
= peptide
+ 5-L-glutamyl amino acid
   Enzyme class 3: E.C.3.4.19.13  - Glutathione hydrolase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Glutathione + H2O = L-cysteinylglycine + L-glutamate
Glutathione
+ H(2)O
= L-cysteinylglycine
+ L-glutamate
Note, where more than one E.C. class is given (as above), each may correspond to a different protein domain or, in the case of polyprotein precursors, to a different mature protein.
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     glutathione metabolic process   1 term 
  Biochemical function     gamma-glutamyltransferase activity     1 term  

 

 
    reference    
 
 
DOI no: 10.1074/jbc.M607490200 J Biol Chem 282:2433-2439 (2007)
PubMed id: 17135273  
 
 
Crystal structure of the gamma-glutamyltranspeptidase precursor protein from Escherichia coli. Structural changes upon autocatalytic processing and implications for the maturation mechanism.
T.Okada, H.Suzuki, K.Wada, H.Kumagai, K.Fukuyama.
 
  ABSTRACT  
 
Gamma-glutamyltranspeptidase (GGT) is an extracellular enzyme that plays a key role in glutathione metabolism. The mature GGT is a heterodimer consisting of L- and S-subunits that is generated by posttranslational cleavage of the peptide bond between Gln-390 and Thr-391 in the precursor protein. Thr-391, which becomes the N-terminal residue of the S-subunit, acts as the active residue in the catalytic reaction. The crystal structure of a mutant GGT, T391A, that is unable to undergo autocatalytic processing, has been determined at 2.55-A resolution. Structural comparison of the precursor protein and mature GGT demonstrates that the structures of the core regions in the two proteins are unchanged, but marked differences are found near the active site. In particular, in the precursor, the segment corresponding to the C-terminal region of the L-subunit occupies the site where the loop (residues 438-449) forms the lid of the gamma-glutamyl group-binding pocket in the mature GGT. This result demonstrates that, upon cleavage of the N-terminal peptide bond of Thr-391, the newly produced C terminus (residues 375-390) flips out, allowing the 438-449 segment to form the gamma-glutamyl group-binding pocket. The electron density map for the T391A protein also identified a water molecule near the carbonyl carbon atom of Gln-390. The spatial arrangement around the water and Thr-391 relative to the scissile peptide bond appears suitable for the initiation of autocatalytic processing, as in other members of the N-terminal nucleophile hydrolase superfamily.
 
  Selected figure(s)  
 
Figure 1.
FIGURE 1. A stereo view of the F[o] – F[c] omit map around the processing site (A molecule). The map was generated on the basis of F[c] calculated from the model, which was derived from the refinement using REFMAC5 (23) omitting residues 385–392 and the water molecule (W4). The map was contoured at the 2.5 level. A ball-and-stick model of the T391A protein is overlaid on the map. The arrow indicates the scissile peptide bond that is cleaved in the wild-type precursor protein (Gln-390 to Thr-391). The figure was prepared using PYMOL (31).
Figure 2.
FIGURE 2. The tertiary structure of the T391A protein. A, a ribbon drawing of the T391A protein (B molecule). The segments in the T391A protein that correspond to the L- and S-subunits are pink and green, respectively, and the P-segment (residue 375–390) is highlighted in orange. Terminal residues that generate invisible segments are labeled. The orange arrow indicates the site at which autocatalytic processing occurs. B, a stereo view of the superimposition of C traces of the T391A protein and mature GGT. The structure of mature GGT (A molecule of SeMet-GGT in (19)) was superimposed on that of the T391A protein (B molecule). P-segment residues in the T391A protein and in mature GGT are orange and blue, respectively. Residues that had C atoms displaced by >1 Å upon processing are in yellow. Residues of mature GGT that are invisible in the T391A protein are shown in black. Regions of invisible residues are circled in green. The distance between Ser-387 C and Thr-391 N in mature GGT is shown. B is rotated by 30° around the vertical axis relative to A. C, a close-up view of the segment Glu-377 to Pro-380. A stick model of mature GGT (blue) is superimposed on the T391A protein (orange). The figures were prepared using PYMOL (31).
 
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (2007, 282, 2433-2439) copyright 2007.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
21298394 I.Castellano, A.Di Salle, A.Merlino, M.Rossi, and F.La Cara (2011).
Gene cloning and protein expression of γ-glutamyltranspeptidases from Thermus thermophilus and Deinococcus radiodurans: comparison of molecular and structural properties with mesophilic counterparts.
  Extremophiles, 15, 259-270.  
20959624 T.Destro, D.Prasad, D.Martignago, I.L.Bernet, A.R.Trentin, I.K.Renu, M.Ferretti, and A.Masi (2011).
Compensatory expression and substrate inducibility of gamma-glutamyl transferase GGT2 isoform in Arabidopsis thaliana.
  J Exp Bot, 62, 805-814.  
19714332 F.H.Hausheer, D.Shanmugarajah, B.D.Leverett, X.Chen, Q.Huang, H.Kochat, P.N.Petluru, and A.R.Parker (2010).
Mechanistic study of BNP7787-mediated cisplatin nephroprotection: modulation of gamma-glutamyl transpeptidase.
  Cancer Chemother Pharmacol, 65, 941-951.  
20673217 H.P.Chang, W.C.Liang, R.C.Lyu, M.C.Chi, T.F.Wang, K.L.Su, H.C.Hung, and L.L.Lin (2010).
Effects of C-terminal truncation on autocatalytic processing of Bacillus licheniformis gamma-glutamyl transpeptidase.
  Biochemistry (Mosc), 75, 919-929.  
20572278 H.Suzuki, C.Yamada, K.Kijima, S.Ishihara, K.Wada, K.Fukuyama, and H.Kumagai (2010).
Enhancement of glutaryl-7-aminocephalosporanic acid acylase activity of gamma-glutamyltranspeptidase of Bacillus subtilis.
  Biotechnol J, 5, 829-837.  
20088880 K.Wada, M.Irie, H.Suzuki, and K.Fukuyama (2010).
Crystal structure of the halotolerant gamma-glutamyltranspeptidase from Bacillus subtilis in complex with glutamate reveals a unique architecture of the solvent-exposed catalytic pocket.
  FEBS J, 277, 1000-1009.
PDB code: 3a75
19340483 R.C.Lyu, H.Y.Hu, L.Y.Kuo, H.F.Lo, P.L.Ong, H.P.Chang, and L.L.Lin (2009).
Role of the conserved Thr399 and Thr417 residues of Bacillus licheniformis gamma-Glutamyltranspeptidase as evaluated by mutational analysis.
  Curr Microbiol, 59, 101-106.  
19535342 R.Wu, S.Richter, R.G.Zhang, V.J.Anderson, D.Missiakas, and A.Joachimiak (2009).
Crystal structure of Bacillus anthracis transpeptidase enzyme CapD.
  J Biol Chem, 284, 24406-24414.
PDB codes: 3g9k 3ga9
18390671 C.Yamada, K.Kijima, S.Ishihara, C.Miwa, K.Wada, T.Okada, K.Fukuyama, H.Kumagai, and H.Suzuki (2008).
Improvement of the glutaryl-7-aminocephalosporanic acid acylase activity of a bacterial gamma-glutamyltranspeptidase.
  Appl Environ Microbiol, 74, 3400-3409.  
18824507 O.D.Ekici, M.Paetzel, and R.E.Dalbey (2008).
Unconventional serine proteases: variations on the catalytic Ser/His/Asp triad configuration.
  Protein Sci, 17, 2023-2037.  
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.