Crystal structures of glutathione- and inhibitor-bound human GGT1: critical interactions within the cysteinylglycine binding site.
crystal structure
crystallography
enzyme
enzyme inhibitor
glutathione
structural biology
γ-glutamyl transferase
γ-glutamyl transpeptidase
Journal
The Journal of biological chemistry
ISSN: 1083-351X
Titre abrégé: J Biol Chem
Pays: United States
ID NLM: 2985121R
Informations de publication
Date de publication:
Historique:
received:
02
10
2020
revised:
06
11
2020
accepted:
13
11
2020
pubmed:
15
11
2020
medline:
15
9
2021
entrez:
14
11
2020
Statut:
ppublish
Résumé
Overexpression of γ-glutamyl transpeptidase (GGT1) has been implicated in an array of human diseases including asthma, reperfusion injury, and cancer. Inhibitors are needed for therapy, but development of potent, specific inhibitors of GGT1 has been hampered by a lack of structural information regarding substrate binding and cleavage. To enhance our understanding of the molecular mechanism of substrate cleavage, we have solved the crystal structures of human GGT1 (hGGT1) with glutathione (a substrate) and a phosphate-glutathione analog (an irreversible inhibitor) bound in the active site. These are the first structures of any eukaryotic GGT with the cysteinylglycine region of the substrate-binding site occupied. These structures and the structure of apo-hGGT reveal movement of amino acid residues within the active site as the substrate binds. Asn-401 and Thr-381 each form hydrogen bonds with two atoms of GSH spanning the γ-glutamyl bond. Three different atoms of hGGT1 interact with the carboxyl oxygen of the cysteine of GSH. Interactions between the enzyme and substrate change as the substrate moves deeper into the active site cleft. The substrate reorients and a new hydrogen bond is formed between the substrate and the oxyanion hole. Thr-381 is locked into a single conformation as an acyl bond forms between the substrate and the enzyme. These data provide insight on a molecular level into the substrate specificity of hGGT1 and provide an explanation for seemingly disparate observations regarding the enzymatic activity of hGGT1 mutants. This knowledge will aid in the design of clinically useful hGGT1 inhibitors.
Identifiants
pubmed: 33187988
pii: S0021-9258(20)00052-6
doi: 10.1074/jbc.RA120.016265
pmc: PMC7949050
pii:
doi:
Substances chimiques
Dipeptides
0
Enzyme Inhibitors
0
cysteinylglycine
384644SZ9T
gamma-Glutamyltransferase
EC 2.3.2.2
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
100066Subventions
Organisme : NIGMS NIH HHS
ID : P41 GM103473
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM125952
Pays : United States
Organisme : NCRR NIH HHS
ID : P41 RR001209
Pays : United States
Organisme : NIGMS NIH HHS
ID : P41 GM111244
Pays : United States
Organisme : NIGMS NIH HHS
ID : P20 GM103640
Pays : United States
Informations de copyright
Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.
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