PDBsum entry 4zcg

Hydrolase

PDB id: 4zcg

Contents

Protein chains

342 a.a.

189 a.a.

Ligands

NAG ×6

GLU

Metals

_NA

_CL ×2

Waters ×428

PDB id:

4zcg

Name:

Hydrolase

Title:

Crystal structure of human ggt1 in complex with glutamate (with all atoms of glutamate)

Structure:

Gamma-glutamyltranspeptidase 1 heavy chain. Chain: a. Fragment: large subunit residues 28-380. Synonym: ggt 1,gamma-glutamyltransferase 1,glutathione hydrolase 1, leukotriene-c4 hydrolase. Engineered: yes. Gamma-glutamyltranspeptidase 1 light chain. Chain: b. Fragment: small subunit residues 381-569.

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: ggt1, ggt. Expressed in: pichia. Expression_system_taxid: 4919.

Resolution:

2.22Å R-factor: 0.148 R-free: 0.185

Authors:

S.Terzyan,M.Hanigan

Key ref:

S.S.Terzyan et al. (2015). Human γ-Glutamyl Transpeptidase 1: STRUCTURES OF THE FREE ENZYME, INHIBITOR-BOUND TETRAHEDRAL TRANSITION STATES, AND GLUTAMATE-BOUND ENZYME REVEAL NOVEL MOVEMENT WITHIN THE ACTIVE SITE DURING CATALYSIS. J Biol Chem, 290, 17576-17586. PubMed id: 26013825 DOI: 10.1074/jbc.M115.659680

Date:

15-Apr-15 Release date: 03-Jun-15

Protein chain

P19440  (GGT1_HUMAN) -  Glutathione hydrolase 1 proenzyme from Homo sapiens

Seq: 569 a.a.

Struc: 342 a.a.*

Protein chain

P19440  (GGT1_HUMAN) -  Glutathione hydrolase 1 proenzyme from Homo sapiens

Seq: 569 a.a.

Struc: 189 a.a.

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class 2: Chains A, B: E.C.2.3.2.2 - gamma-glutamyltransferase.
• Reaction: an N-terminal (5-L-glutamyl)-[peptide] + an alpha-amino acid = 5-L- glutamyl amino acid + an N-terminal L-alpha-aminoacyl-[peptide]

N-terminal (5-L-glutamyl)-[peptide] (Bound ligand (Het Group name = GLU) matches with 45.45% similarity) + alpha-amino acid = 5-L- glutamyl amino acid (Bound ligand (Het Group name = NAG) matches with 45.00% similarity) + N-terminal L-alpha-aminoacyl-[peptide]

• Enzyme class 3: Chains A, B: E.C.3.4.19.13 - glutathione gamma-glutamate hydrolase.
• Reaction:
  1. glutathione + H2O = L-cysteinylglycine + L-glutamate
  2. an S-substituted glutathione + H2O = an S-substituted L-cysteinylglycine + L-glutamate

glutathione + H2O = L-cysteinylglycine + L-glutamate (Bound ligand (Het Group name = GLU) corresponds exactly)

S-substituted glutathione + H2O = S-substituted L-cysteinylglycine + L-glutamate (Bound ligand (Het Group name = GLU) corresponds exactly)

• Enzyme class 4: Chains A, B: E.C.3.4.19.14 - leukotriene-C4 hydrolase.
• Reaction: leukotriene C4 + H2O = leukotriene D4 + L-glutamate

leukotriene C4 + H2O = leukotriene D4 (Bound ligand (Het Group name = GLU) corresponds exactly) + 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

reference

DOI no: 10.1074/jbc.M115.659680

J Biol Chem 290:17576-17586 (2015)

PubMed id: 26013825

Human γ-Glutamyl Transpeptidase 1: STRUCTURES OF THE FREE ENZYME, INHIBITOR-BOUND TETRAHEDRAL TRANSITION STATES, AND GLUTAMATE-BOUND ENZYME REVEAL NOVEL MOVEMENT WITHIN THE ACTIVE SITE DURING CATALYSIS.

S.S.Terzyan, A.W.Burgett, A.Heroux, C.A.Smith, B.H.Mooers, M.H.Hanigan.

ABSTRACT

γ-Glutamyl transpeptidase 1 (GGT1) is a cell surface, N-terminal nucleophile hydrolase that cleaves glutathione and other γ-glutamyl compounds. GGT1 expression is essential in cysteine homeostasis, and its induction has been implicated in the pathology of asthma, reperfusion injury, and cancer. In this study, we report four new crystal structures of human GGT1 (hGGT1) that show conformational changes within the active site as the enzyme progresses from the free enzyme to inhibitor-bound tetrahedral transition states and finally to the glutamate-bound structure prior to the release of this final product of the reaction. The structure of the apoenzyme shows flexibility within the active site. The serine-borate-bound hGGT1 crystal structure demonstrates that serine-borate occupies the active site of the enzyme, resulting in an enzyme-inhibitor complex that replicates the enzyme's tetrahedral intermediate/transition state. The structure of GGsTop-bound hGGT1 reveals its interactions with the enzyme and why neutral phosphonate diesters are more potent inhibitors than monoanionic phosphonates. These structures are the first structures for any eukaryotic GGT that include a molecule in the active site covalently bound to the catalytic Thr-381. The glutamate-bound structure shows the conformation of the enzyme prior to release of the final product and reveals novel information regarding the displacement of the main chain atoms that form the oxyanion hole and movement of the lid loop region when the active site is occupied. These data provide new insights into the mechanism of hGGT1-catalyzed reactions and will be invaluable in the development of new classes of hGGT1 inhibitors for therapeutic use.