skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Discovery of an Inhibitor of Z-Alpha1 Antitrypsin Polymerization

Journal Article · · PLoS ONE
 [1];  [2];  [1];  [3];
  1. 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
  2. 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
  3. 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

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.

Research Organization:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Organization:
USDOE
Grant/Contract Number:
AC05-00OR22725
OSTI ID:
1334442
Journal Information:
PLoS ONE, Vol. 10, Issue 5; ISSN 1932-6203
Publisher:
Public Library of ScienceCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 7 works
Citation information provided by
Web of Science

References (40)

Alpha-1-Antitrypsin Deficiency: Importance of Proteasomal and Autophagic Degradative Pathways in Disposal of Liver Disease–Associated Protein Aggregates journal February 2011
Serpinopathies and the conformational dementias journal October 2002
Preventing serpin aggregation: The molecular mechanism of citrate action upon antitrypsin unfolding journal December 2008
Structure of a serpin–protease complex shows inhibition by deformation journal October 2000
Polymers of α 1 -Antitrypsin Are Chemotactic for Human Neutrophils: A New Paradigm for the Pathogenesis of Emphysema journal June 2002
Small Molecules Block the Polymerization of Z α 1 -Antitrypsin and Increase the Clearance of Intracellular Aggregates journal November 2007
Inactive conformation of the serpin alpha 1-antichymotrypsin indicates two-stage insertion of the reactive loop: Implications for inhibitory function and conformational disease journal January 2000
Molecular basis of alpha-1-antitrypsin deficiency journal June 1988
6-mer Peptide Selectively Anneals to a Pathogenic Serpin Conformation and Blocks Polymerization: IMPLICATIONS FOR THE PREVENTION OF Z α 1 -ANTITRYPSIN-RELATED CIRRHOSIS journal December 2001
α 1 -Antitrypsin Polymerization:  A Fluorescence Correlation Spectroscopic Study journal February 2005
Crystal structure of a stable dimer reveals the molecular basis of serpin polymerization journal October 2008
Chemical chaperones mediate increased secretion of mutant alpha 1-antitrypsin (alpha 1-AT) Z: A potential pharmacological strategy for prevention of liver injury and emphysema in alpha 1-AT deficiency journal February 2000
A Kinetic Mechanism for the Polymerization of α 1 -Antitrypsin journal April 1999
Lack of Effect of Oral 4-Phenylbutyrate on Serum Alpha-1-Antitrypsin in Patients with α-1-Antitrypsin Deficiency: A Preliminary Study journal January 2004
Ensemble-Based Computational Approach Discriminates Functional Activity of p53 Cancer and Rescue Mutants journal October 2011
Topography of a 2.0 Å structure of α 1 -antitrypsin reveals targets for rational drug design to prevent conformational disease journal January 2000
A novel monoclonal antibody to characterize pathogenic polymers in liver disease associated with α 1 -antitrypsin deficiency journal May 2010
Lung Polymers in Z α 1 -Antitrypsin Deficiency-related Emphysema journal May 1998
Defining the mechanism of polymerization in the serpinopathies journal September 2010
The generalized Born/volume integral implicit solvent model: Estimation of the free energy of hydration using London dispersion instead of atomic surface area journal February 2008
Molecular basis of α 1 ‐antitrypsin deficiency revealed by the structure of a domain‐swapped trimer journal October 2011
Prevention of Polymerization of M and Z α 1 -Antitrypsin ( α 1 -AT) with Trimethylamine N -Oxide: Implications for the Treatment of α 1 -AT Deficiency journal June 2001
Protonate3D: Assignment of ionization states and hydrogen coordinates to macromolecular structures journal September 2008
Quantitative isolation of ?1AT mutant Z protein polymers from human and mouse livers and the effect of heat journal January 2005
The mechanism of Z α1-antitrypsin accumulation in the liver journal June 1992
Loop-sheet polymerization: the mechanism of alpha1-antitrypsin deficiency journal August 2000
α1-antitrypsin deficiency and liver disease: Clinical presentation, diagnosis and treatment journal July 1991
Targeting a Surface Cavity of α 1 -Antitrypsin to Prevent Conformational Disease journal June 2003
Crystallographic and Cellular Characterisation of Two Mechanisms Stabilising the Native Fold of α1-Antitrypsin: Implications for Disease and Drug Design journal April 2009
Ensemble-Based Virtual Screening Reveals Potential Novel Antiviral Compounds for Avian Influenza Neuraminidase journal July 2008
Molecular gymnastics: serpin structure, folding and misfolding journal December 2006
Prevalence of α1-antitrypsin deficiency alleles PI*S and PI*Z worldwide and effective screening for each of the five phenotypic classes PI*MS, PI*MZ, PI*SS, PI*SZ, and PI*ZZ: a comprehensive review journal July 2012
The molecular and cellular pathology of α1-antitrypsin deficiency journal February 2014
Intravenous alpha-1 antitrypsin augmentation therapy for treating patients with alpha-1 antitrypsin deficiency and lung disease journal September 2016
Preventing and reversing the cellular consequences of Z alpha-1 antitrypsin accumulation by targeting s4A journal July 2012
Polymers of Z α1-Antitrypsin Co-Localize with Neutrophils in Emphysematous Alveoli and Are Chemotactic in Vivo journal February 2005
Chemical beauty contest journal January 2012
Targeting a Surface Cavity of α1-Antitrypsin to Prevent Conformational Disease journal April 2003
Risk of Cirrhosis and Primary Liver Cancer in Alpha 1 -Antitrypsin Deficiency journal March 1986
Z-type alpha 1-antitrypsin is less competent than M1-type alpha 1-antitrypsin as an inhibitor of neutrophil elastase. journal November 1987

Cited By (1)

A small molecule chaperone rescues the stability and activity of a cancer‐associated variant of NAD(P)H:quinone oxidoreductase 1 in vitro journal October 2019