Discovery of an Inhibitor of Z-Alpha1 Antitrypsin Polymerization
Abstract
Polymerization of the Z variant alpha-1-antitrypsin (Z-α1AT) results in the most common and severe form of α1AT deficiency (α1ATD), a debilitating genetic disorder whose clinical manifestations range from asymptomatic to fatal liver and/or lung disease. As the altered conformation of Z-α1AT and its attendant aggregation are responsible for pathogenesis, the polymerization process per se has become a major target for the development of therapeutics. Based on the ability of Z-alpha 1AT to aggregate by recruiting the reactive center loop (RCL) of another Z-α1AT into its s4A cavity, we developed a high-throughput screening assay that uses a modified 6-mer peptide mimicking the RCL to screen for inhibitors of Z-α1AT polymer growth. We used a subset of compounds from the Library of Pharmacologically Active Compounds (LOPAC) with molecular weights ranging from 300 to 700 Da, to evaluate the assay's capabilities. The inhibitor S-(4-nitrobenzyl)-6-thioguanosine was identified as a lead compound and its ability to prevent Z-α1AT polymerization confirmed by secondary assays. In order to further investigate the binding location of S-(4-nitrobenzyl)-6-thioguanosine, an in silico strategy was pursued and the intermediate alpha 1AT M* state modeled to allow molecular docking simulations and explore various potential binding sites. Docking results predict that S-(4-nitrobenzyl)-6-thioguanosine can bindmore »
- Authors:
-
- Univ. of Tennessee Health Science Center, Knoxville, TN (United States). Dept. of Medicine; Univ. of Tennessee, Knoxville, TN (United States). Gradate School of Genome Science and Technology
- Univ. of Tennessee, Knoxville, TN (United States). Gradate School of Genome Science and Technology; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Center for Molecular Biophysics
- Univ. of Tennessee, Knoxville, TN (United States). Gradate School of Genome Science and Technology and Dept. of Biochemistry and Cellular and Molecular Biology; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Center for Molecular Biophysics
- Publication Date:
- Research Org.:
- Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1334442
- Grant/Contract Number:
- AC05-00OR22725
- Resource Type:
- Accepted Manuscript
- Journal Name:
- PLoS ONE
- Additional Journal Information:
- Journal Volume: 10; Journal Issue: 5; Journal ID: ISSN 1932-6203
- Publisher:
- Public Library of Science
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Berthelier, Valerie, Harris, Jason Brett, Estenson, Kasey Noel, Baudry, Jerome, and Carloni, Paolo. Discovery of an Inhibitor of Z-Alpha1 Antitrypsin Polymerization. United States: N. p., 2015.
Web. doi:10.1371/journal.pone.0126256.
Berthelier, Valerie, Harris, Jason Brett, Estenson, Kasey Noel, Baudry, Jerome, & Carloni, Paolo. Discovery of an Inhibitor of Z-Alpha1 Antitrypsin Polymerization. United States. https://doi.org/10.1371/journal.pone.0126256
Berthelier, Valerie, Harris, Jason Brett, Estenson, Kasey Noel, Baudry, Jerome, and Carloni, Paolo. Mon .
"Discovery of an Inhibitor of Z-Alpha1 Antitrypsin Polymerization". United States. https://doi.org/10.1371/journal.pone.0126256. https://www.osti.gov/servlets/purl/1334442.
@article{osti_1334442,
title = {Discovery of an Inhibitor of Z-Alpha1 Antitrypsin Polymerization},
author = {Berthelier, Valerie and Harris, Jason Brett and Estenson, Kasey Noel and Baudry, Jerome and Carloni, Paolo},
abstractNote = {Polymerization of the Z variant alpha-1-antitrypsin (Z-α1AT) results in the most common and severe form of α1AT deficiency (α1ATD), a debilitating genetic disorder whose clinical manifestations range from asymptomatic to fatal liver and/or lung disease. As the altered conformation of Z-α1AT and its attendant aggregation are responsible for pathogenesis, the polymerization process per se has become a major target for the development of therapeutics. Based on the ability of Z-alpha 1AT to aggregate by recruiting the reactive center loop (RCL) of another Z-α1AT into its s4A cavity, we developed a high-throughput screening assay that uses a modified 6-mer peptide mimicking the RCL to screen for inhibitors of Z-α1AT polymer growth. We used a subset of compounds from the Library of Pharmacologically Active Compounds (LOPAC) with molecular weights ranging from 300 to 700 Da, to evaluate the assay's capabilities. The inhibitor S-(4-nitrobenzyl)-6-thioguanosine was identified as a lead compound and its ability to prevent Z-α1AT polymerization confirmed by secondary assays. In order to further investigate the binding location of S-(4-nitrobenzyl)-6-thioguanosine, an in silico strategy was pursued and the intermediate alpha 1AT M* state modeled to allow molecular docking simulations and explore various potential binding sites. Docking results predict that S-(4-nitrobenzyl)-6-thioguanosine can bind at the s4A cavity and at the edge of beta-sheet A. The former binding site would directly block RCL insertion whereas the latter site would prevent beta-sheet A from expanding between s3A/s5A, and thus indirectly impede RCL insertion. Our investigations have revealed a novel compound that inhibits the formation of Z-α1AT polymers, as well as in vitro and in silico strategies for identifying and characterizing additional blocking molecules of Z-α1AT polymerization.},
doi = {10.1371/journal.pone.0126256},
journal = {PLoS ONE},
number = 5,
volume = 10,
place = {United States},
year = {Mon May 11 00:00:00 EDT 2015},
month = {Mon May 11 00:00:00 EDT 2015}
}
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