The Importance of N186 in the Alpha-1-Antitrypsin Shutter Region Is Revealed by the Novel Bologna Deficiency Variant
Alpha-1-antitrypsin (AAT) deficiency causes pulmonary disease due to decreased levels of circulating AAT and consequently unbalanced protease activity in the lungs. Deposition of specific AAT variants, such as the common Z AAT, within hepatocytes may also result in liver disease. These deposits are...
Main Authors: | , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2021-05-01
|
Series: | International Journal of Molecular Sciences |
Subjects: | |
Online Access: | https://www.mdpi.com/1422-0067/22/11/5668 |
id |
doaj-60628c5796ca46fe8964acc684f1df3e |
---|---|
record_format |
Article |
spelling |
doaj-60628c5796ca46fe8964acc684f1df3e2021-06-01T01:12:35ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-05-01225668566810.3390/ijms22115668The Importance of N186 in the Alpha-1-Antitrypsin Shutter Region Is Revealed by the Novel Bologna Deficiency VariantRiccardo Ronzoni0Ilaria Ferrarotti1Emanuela D’Acunto2Alice M. Balderacchi3Stefania Ottaviani4David A. Lomas5James A. Irving6Elena Miranda7Annamaria Fra8UCL Respiratory and the Institute of Structural and Molecular Biology, University College London, London WC1E 6JF, UKPneumology Unit, Centre for Diagnosis of Inherited Alpha-1 Antitrypsin Deficiency, Department of Internal Medicine and Therapeutics, IRCCS San Matteo Hospital Foundation, University of Pavia, 27100 Pavia, ItalyDepartment of Biology and Biotechnologies ‘Charles Darwin’, Sapienza University of Rome, 00185 Rome, ItalyPneumology Unit, Centre for Diagnosis of Inherited Alpha-1 Antitrypsin Deficiency, Department of Internal Medicine and Therapeutics, IRCCS San Matteo Hospital Foundation, University of Pavia, 27100 Pavia, ItalyPneumology Unit, Centre for Diagnosis of Inherited Alpha-1 Antitrypsin Deficiency, Department of Internal Medicine and Therapeutics, IRCCS San Matteo Hospital Foundation, University of Pavia, 27100 Pavia, ItalyUCL Respiratory and the Institute of Structural and Molecular Biology, University College London, London WC1E 6JF, UKUCL Respiratory and the Institute of Structural and Molecular Biology, University College London, London WC1E 6JF, UKDepartment of Biology and Biotechnologies ‘Charles Darwin’, Sapienza University of Rome, 00185 Rome, ItalyDepartment of Molecular and Translational Medicine, University of Brescia, viale Europa 11, 25123 Brescia, ItalyAlpha-1-antitrypsin (AAT) deficiency causes pulmonary disease due to decreased levels of circulating AAT and consequently unbalanced protease activity in the lungs. Deposition of specific AAT variants, such as the common Z AAT, within hepatocytes may also result in liver disease. These deposits are comprised of ordered polymers of AAT formed by an inter-molecular domain swap. The discovery and characterization of rare variants of AAT and other serpins have historically played a crucial role in the dissection of the structural mechanisms leading to AAT polymer formation. Here, we report a severely deficient shutter region variant, Bologna AAT (N186Y), which was identified in five unrelated subjects with different geographical origins. We characterized the new variant by expression in cellular models in comparison with known polymerogenic AAT variants. Bologna AAT showed secretion deficiency and intracellular accumulation as detergent-insoluble polymers. Extracellular polymers were detected in both the culture media of cells expressing Bologna AAT and in the plasma of a patient homozygous for this variant. Structural modelling revealed that the mutation disrupts the hydrogen bonding network in the AAT shutter region. These data support a crucial coordinating role for asparagine 186 and the importance of this network in promoting formation of the native structure.https://www.mdpi.com/1422-0067/22/11/5668liver storage diseasealpha-1-antitrypsin deficiencyendoplasmic reticulumprotein aggregation<i>SERPINA1</i> allelesserpinopathies |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Riccardo Ronzoni Ilaria Ferrarotti Emanuela D’Acunto Alice M. Balderacchi Stefania Ottaviani David A. Lomas James A. Irving Elena Miranda Annamaria Fra |
spellingShingle |
Riccardo Ronzoni Ilaria Ferrarotti Emanuela D’Acunto Alice M. Balderacchi Stefania Ottaviani David A. Lomas James A. Irving Elena Miranda Annamaria Fra The Importance of N186 in the Alpha-1-Antitrypsin Shutter Region Is Revealed by the Novel Bologna Deficiency Variant International Journal of Molecular Sciences liver storage disease alpha-1-antitrypsin deficiency endoplasmic reticulum protein aggregation <i>SERPINA1</i> alleles serpinopathies |
author_facet |
Riccardo Ronzoni Ilaria Ferrarotti Emanuela D’Acunto Alice M. Balderacchi Stefania Ottaviani David A. Lomas James A. Irving Elena Miranda Annamaria Fra |
author_sort |
Riccardo Ronzoni |
title |
The Importance of N186 in the Alpha-1-Antitrypsin Shutter Region Is Revealed by the Novel Bologna Deficiency Variant |
title_short |
The Importance of N186 in the Alpha-1-Antitrypsin Shutter Region Is Revealed by the Novel Bologna Deficiency Variant |
title_full |
The Importance of N186 in the Alpha-1-Antitrypsin Shutter Region Is Revealed by the Novel Bologna Deficiency Variant |
title_fullStr |
The Importance of N186 in the Alpha-1-Antitrypsin Shutter Region Is Revealed by the Novel Bologna Deficiency Variant |
title_full_unstemmed |
The Importance of N186 in the Alpha-1-Antitrypsin Shutter Region Is Revealed by the Novel Bologna Deficiency Variant |
title_sort |
importance of n186 in the alpha-1-antitrypsin shutter region is revealed by the novel bologna deficiency variant |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2021-05-01 |
description |
Alpha-1-antitrypsin (AAT) deficiency causes pulmonary disease due to decreased levels of circulating AAT and consequently unbalanced protease activity in the lungs. Deposition of specific AAT variants, such as the common Z AAT, within hepatocytes may also result in liver disease. These deposits are comprised of ordered polymers of AAT formed by an inter-molecular domain swap. The discovery and characterization of rare variants of AAT and other serpins have historically played a crucial role in the dissection of the structural mechanisms leading to AAT polymer formation. Here, we report a severely deficient shutter region variant, Bologna AAT (N186Y), which was identified in five unrelated subjects with different geographical origins. We characterized the new variant by expression in cellular models in comparison with known polymerogenic AAT variants. Bologna AAT showed secretion deficiency and intracellular accumulation as detergent-insoluble polymers. Extracellular polymers were detected in both the culture media of cells expressing Bologna AAT and in the plasma of a patient homozygous for this variant. Structural modelling revealed that the mutation disrupts the hydrogen bonding network in the AAT shutter region. These data support a crucial coordinating role for asparagine 186 and the importance of this network in promoting formation of the native structure. |
topic |
liver storage disease alpha-1-antitrypsin deficiency endoplasmic reticulum protein aggregation <i>SERPINA1</i> alleles serpinopathies |
url |
https://www.mdpi.com/1422-0067/22/11/5668 |
work_keys_str_mv |
AT riccardoronzoni theimportanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT ilariaferrarotti theimportanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT emanueladacunto theimportanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT alicembalderacchi theimportanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT stefaniaottaviani theimportanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT davidalomas theimportanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT jamesairving theimportanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT elenamiranda theimportanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT annamariafra theimportanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT riccardoronzoni importanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT ilariaferrarotti importanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT emanueladacunto importanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT alicembalderacchi importanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT stefaniaottaviani importanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT davidalomas importanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT jamesairving importanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT elenamiranda importanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant AT annamariafra importanceofn186inthealpha1antitrypsinshutterregionisrevealedbythenovelbolognadeficiencyvariant |
_version_ |
1721412868185784320 |