DB code: D00192

RLCP classification 1.13.11400.1261 : Hydrolysis
CATH domain 3.40.630.10 : Aminopeptidase Catalytic domain
3.30.70.360 : Alpha-Beta Plaits
E.C. 3.4.17.11
CSA 1cg2
M-CSA 1cg2
MACiE

CATH domain Related DB codes (homologues)
3.40.630.10 : Aminopeptidase D00512 S00406 S00407 D00193 D00467

Uniprot Enzyme Name
UniprotKB Protein name Synonyms MEROPS Pfam
P06621 Carboxypeptidase G2 (CPDG2) (EC 3.4.17.11) (Folate hydrolase G2) (Glutamate carboxypeptidase) (Pteroylmonoglutamic acid hydrolase G2)AltName: INN=Glucarpidase;
None M20.001 (Metallo)
PF07687 (M20_dimer)
PF01546 (Peptidase_M20)
[Graphical View]

KEGG enzyme name
glutamate carboxypeptidase
carboxypeptidase G
carboxypeptidase G1
carboxypeptidase G2
glutamyl carboxypeptidase
N-pteroyl-L-glutamate hydrolase

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
P06621 CBPG_PSES6 Release of C-terminal glutamate residues from a wide range of N-acylating moieties, including peptidyl, aminoacyl, benzoyl, benzyloxycarbonyl, folyl and pteroyl groups. Homodimer. Binds 2 zinc ions per subunit.

KEGG Pathways
Map code Pathways E.C.

Compound table
Cofactors Substrates Products Intermediates
KEGG-id C00038 C00001 C00012 C00504 C00012 C00921 C00025
E.C.
Compound Zinc H2O Peptide Folic acid Peptide Dihydropteroate L-Glutamate
Type heavy metal H2O peptide/protein amino acids,amide group,amine group,aromatic ring (only carbon atom),aromatic ring (with nitrogen atoms),carboxyl group peptide/protein amide group,amine group,aromatic ring (only carbon atom),aromatic ring (with nitrogen atoms),carboxyl group amino acids,carboxyl group
ChEBI 29105
15377
27470
4581
16015
PubChem 32051
22247451
962
6037
170
33032
44272391
88747398
1cg2A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:2x_ZN Unbound Unbound Unbound Unbound Unbound
1cg2B01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:2x_ZN Unbound Unbound Unbound Unbound Unbound
1cg2C01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:2x_ZN Unbound Unbound Unbound Unbound Unbound
1cg2D01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:2x_ZN Unbound Unbound Unbound Unbound Unbound
1cg2A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:_ZN Unbound Unbound Unbound Unbound Unbound
1cg2B02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:_ZN Unbound Unbound Unbound Unbound Unbound
1cg2C02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:_ZN Unbound Unbound Unbound Unbound Unbound
1cg2D02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:_ZN Unbound Unbound Unbound Unbound Unbound

Reference for Active-site residues
resource references E.C.
Swiss-prot;P06621 & literature [3]

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
1cg2A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 175 GLU 176;HIS 385(Zinc-1 binding);HIS 112;GLU 200(Zinc-2 binding);ASP 141(Zinc-1 and -2 binding)
1cg2B01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 175 GLU 176;HIS 385(Zinc-1 binding);HIS 112;GLU 200(Zinc-2 binding);ASP 141(Zinc-1 and -2 binding)
1cg2C01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 175 GLU 176;HIS 385(Zinc-1 binding);HIS 112;GLU 200(Zinc-2 binding);ASP 141(Zinc-1 and -2 binding)
1cg2D01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 175 GLU 176;HIS 385(Zinc-1 binding);HIS 112;GLU 200(Zinc-2 binding);ASP 141(Zinc-1 and -2 binding)
1cg2A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain
1cg2B02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain
1cg2C02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain
1cg2D02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain

References for Catalytic Mechanism
References Sections No. of steps in catalysis
[3]
p.342-344

References
[1]
Resource
Comments
Medline ID
PubMed ID 2067015
Journal J Mol Biol
Year 1991
Volume 220
Pages 17-8
Authors Lloyd LF, Collyer CA, Sherwood RF
Title Crystallization and preliminary crystallographic analysis of carboxypeptidase G2 from Pseudomonas sp. strain RS-16.
Related PDB
Related UniProtKB
[2]
Resource
Comments
Medline ID
PubMed ID 9187245
Journal Biochim Biophys Acta
Year 1997
Volume 1339
Pages 247-52
Authors Rawlings ND, Barrett AJ
Title Structure of membrane glutamate carboxypeptidase.
Related PDB
Related UniProtKB
[3]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS)
Medline ID 97238930
PubMed ID 9083113
Journal Structure
Year 1997
Volume 5
Pages 337-47
Authors Rowsell S, Pauptit RA, Tucker AD, Melton RG, Blow DM, Brick P
Title Crystal structure of carboxypeptidase G2, a bacterial enzyme with applications in cancer therapy.
Related PDB 1cg2
Related UniProtKB P06621
[4]
Resource
Comments
Medline ID
PubMed ID 9666335
Journal Curr Opin Struct Biol
Year 1998
Volume 8
Pages 380-7
Authors Murzin AG
Title How far divergent evolution goes in proteins.
Related PDB
Related UniProtKB
[5]
Resource
Comments
Medline ID
PubMed ID 10360747
Journal Bioorg Med Chem Lett
Year 1999
Volume 9
Pages 1415-8
Authors Rodriguez CE, Lu H, Dinh TT, Mlodnosky KL, Dastgah A, Lam VQ, Nichols CB, Berkman CE
Title Competitive inhibition of a glutamate carboxypeptidase by phosphonamidothionate derivatives of glutamic acid.
Related PDB
Related UniProtKB
[6]
Resource
Comments
Medline ID
PubMed ID 10090777
Journal J Med Chem
Year 1999
Volume 42
Pages 951-6
Authors Khan TH, Eno-Amooquaye EA, Searle F, Browne PJ, Osborn HM, Burke PJ
Title Novel inhibitors of carboxypeptidase G2 (CPG2): potential use in antibody-directed enzyme prodrug therapy.
Related PDB
Related UniProtKB
[7]
Resource
Comments
Medline ID
PubMed ID 9882712
Journal Mol Pharmacol
Year 1999
Volume 55
Pages 179-85
Authors Speno HS, Luthi-Carter R, Macias WL, Valentine SL, Joshi AR, Coyle JT
Title Site-directed mutagenesis of predicted active site residues in glutamate carboxypeptidase II.
Related PDB
Related UniProtKB
[8]
Resource
Comments
Medline ID
PubMed ID 10595564
Journal Protein Sci
Year 1999
Volume 8
Pages 2546-9
Authors Mahadevan D, Saldanha JW
Title The extracellular regions of PSMA and the transferrin receptor contain an aminopeptidase domain: implications for drug design.
Related PDB
Related UniProtKB
[9]
Resource
Comments
Medline ID
PubMed ID 10531064
Journal Science
Year 1999
Volume 286
Pages 779-82
Authors Lawrence CM, Ray S, Babyonyshev M, Galluser R, Borhani DW, Harrison SC
Title Crystal structure of the ectodomain of human transferrin receptor.
Related PDB
Related UniProtKB
[10]
Resource
Comments
Medline ID
PubMed ID 10715144
Journal J Med Chem
Year 2000
Volume 43
Pages 772-4
Authors Nan F, Bzdega T, Pshenichkin S, Wroblewski JT, Wroblewska B, Neale JH, Kozikowski AP
Title Dual function glutamate-related ligands: discovery of a novel, potent inhibitor of glutamate carboxypeptidase II possessing mGluR3 agonist activity.
Related PDB
Related UniProtKB
[11]
Resource
Comments
Medline ID
PubMed ID 10739875
Journal Pharmacol Ther
Year 2000
Volume 85
Pages 207-15
Authors Galivan J, Ryan TJ, Chave K, Rhee M, Yao R, Yin D
Title Glutamyl hydrolase. pharmacological role and enzymatic characterization.
Related PDB
Related UniProtKB
[12]
Resource
Comments
Medline ID
PubMed ID 11249132
Journal Bioorg Med Chem
Year 2001
Volume 9
Pages 395-402
Authors Lu H, Berkman CE
Title Stereoselective inhibition of glutamate carboxypeptidase by chiral phosphonothioic acids.
Related PDB
Related UniProtKB
[13]
Resource
Comments
Medline ID
PubMed ID 11375762
Journal Curr Med Chem
Year 2001
Volume 8
Pages 949-57
Authors Jackson PF, Slusher BS
Title Design of NAALADase inhibitors: a novel neuroprotective strategy.
Related PDB
Related UniProtKB
[14]
Resource
Comments
Medline ID
PubMed ID 11708918
Journal J Med Chem
Year 2001
Volume 44
Pages 4170-5
Authors Jackson PF, Tays KL, Maclin KM, Ko YS, Li W, Vitharana D, Tsukamoto T, Stoermer D, Lu XC, Wozniak K, Slusher BS
Title Design and pharmacological activity of phosphinic acid based NAALADase inhibitors.
Related PDB
Related UniProtKB
[15]
Resource
Comments
Medline ID
PubMed ID 11462970
Journal J Med Chem
Year 2001
Volume 44
Pages 298-301
Authors Kozikowski AP, Nan F, Conti P, Zhang J, Ramadan E, Bzdega T, Wroblewska B, Neale JH, Pshenichkin S, Wroblewski JT
Title Design of remarkably simple, yet potent urea-based inhibitors of glutamate carboxypeptidase II (NAALADase).
Related PDB
Related UniProtKB
[16]
Resource
Comments
Medline ID
PubMed ID 12127534
Journal Bioorg Med Chem Lett
Year 2002
Volume 12
Pages 2189-92
Authors Tsukamoto T, Flanary JM, Rojas C, Slusher BS, Valiaeva N, Coward JK
Title Phosphonate and phosphinate analogues of N-acylated gamma-glutamylglutamate. potent inhibitors of glutamate carboxypeptidase II.
Related PDB
Related UniProtKB
[17]
Resource
Comments
Medline ID
PubMed ID 11856302
Journal Eur J Biochem
Year 2002
Volume 269
Pages 443-50
Authors Hakansson K, Miller CG
Title Structure of peptidase T from Salmonella typhimurium.
Related PDB
Related UniProtKB
[18]
Resource
Comments
Medline ID
PubMed ID 12213057
Journal J Med Chem
Year 2002
Volume 45
Pages 4140-52
Authors Rong SB, Zhang J, Neale JH, Wroblewski JT, Wang S, Kozikowski AP
Title Molecular modeling of the interactions of glutamate carboxypeptidase II with its potent NAAG-based inhibitors.
Related PDB
Related UniProtKB
[19]
Resource
Comments
Medline ID
PubMed ID 11921443
Journal Proteomics
Year 2002
Volume 2
Pages 271-9
Authors Spencer DI, Robson L, Purdy D, Whitelegg NR, Michael NP, Bhatia J, Sharma SK, Rees AR, Minton NP, Begent RH, Chester KA
Title A strategy for mapping and neutralizing conformational immunogenic sites on protein therapeutics.
Related PDB
Related UniProtKB
[20]
Resource
Comments
Medline ID
PubMed ID 12885660
Journal Biophys J
Year 2003
Volume 85
Pages 1165-75
Authors Occhipinti E, Martelli PL, Spinozzi F, Corsi F, Formantici C, Molteni L, Amenitsch H, Mariani P, Tortora P, Casadio R
Title 3D structure of Sulfolobus solfataricus carboxypeptidase developed by molecular modeling is confirmed by site-directed mutagenesis and small angle X-ray scattering.
Related PDB
Related UniProtKB
[21]
Resource
Comments
Medline ID
PubMed ID 15027864
Journal J Med Chem
Year 2004
Volume 47
Pages 1729-38
Authors Kozikowski AP, Zhang J, Nan F, Petukhov PA, Grajkowska E, Wroblewski JT, Yamamoto T, Bzdega T, Wroblewska B, Neale JH
Title Synthesis of urea-based inhibitors as active site probes of glutamate carboxypeptidase II: efficacy as analgesic agents.
Related PDB
Related UniProtKB

Comments
This enzyme belongs to the peptidase family-M20A.
According to the literature [3], this enzyme must have a similar mechanism to that of bacterial leucine aminopeptidase (S00406 in this database). The catalytic reaction proceeds as follows (see [3]);
(1) Glu175 acts as a general base, which activate the bridging water between the two catalytic zinc ions.
(2) The activated bridging water molecule acts as a nucleophile to hydrolyze the scissile peptide bond.

Created Updated
2004-08-19 2009-02-26