5B5T

Crystal Structure of Escherichia coli Gamma-Glutamyltranspeptidase in Complex with peptidyl phosphonate inhibitor 1b

Summary for 5B5T

• Entry DOI: 10.2210/pdb5b5t/pdb
• Descriptor: Gamma-glutamyltranspeptidase large chain, Gamma-glutamyltranspeptidase small chain, (2~{S})-2-azanyl-4-[(2~{R})-1-(2-hydroxy-2-oxoethylamino)-1-oxidanylidene-butan-2-yl]oxyphosphonoyl-butanoic acid, ... (5 entities in total)
• Functional Keywords: glutathione, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor
• Biological source: Escherichia coli (strain K12); More
• Total number of polymer chains: 4
• Total formula weight: 119380.05

Authors

Wada, K.,Fukuyama, K. (deposition date: 2016-05-18, release date: 2016-09-28, Last modification date: 2023-11-08)

Primary citation

Kamiyama, A.,Nakajima, M.,Han, L.,Wada, K.,Mizutani, M.,Tabuchi, Y.,Kojima-Yuasa, A.,Matsui-Yuasa, I.,Suzuki, H.,Fukuyama, K.,Watanabe, B.,Hiratake, J.
Phosphonate-based irreversible inhibitors of human gamma-glutamyl transpeptidase (GGT). GGsTop is a non-toxic and highly selective inhibitor with critical electrostatic interaction with an active-site residue Lys562 for enhanced inhibitory activity
Bioorg.Med.Chem., 24:5340-5352, 2016

PubMed Abstract: γ-Glutamyl transpeptidase (GGT, EC 2.3.2.2) that catalyzes the hydrolysis and transpeptidation of glutathione and its S-conjugates is involved in a number of physiological and pathological processes through glutathione metabolism and is an attractive pharmaceutical target. We report here the evaluation of a phosphonate-based irreversible inhibitor, 2-amino-4-{[3-(carboxymethyl)phenoxy](methoyl)phosphoryl}butanoic acid (GGsTop) and its analogues as a mechanism-based inhibitor of human GGT. GGsTop is a stable compound, but inactivated the human enzyme significantly faster than the other phosphonates, and importantly did not inhibit a glutamine amidotransferase. The structure-activity relationships, X-ray crystallography with Escherichia coli GGT, sequence alignment and site-directed mutagenesis of human GGT revealed a critical electrostatic interaction between the terminal carboxylate of GGsTop and the active-site residue Lys562 of human GGT for potent inhibition. GGsTop showed no cytotoxicity toward human fibroblasts and hepatic stellate cells up to 1mM. GGsTop serves as a non-toxic, selective and highly potent irreversible GGT inhibitor that could be used for various in vivo as well as in vitro biochemical studies.

PubMed: 27622749
DOI: 10.1016/j.bmc.2016.08.050

Experimental method

X-RAY DIFFRACTION (1.7 Å)