Point Mutation of a Non-Elastase-Binding Site in Human α1-Antitrypsin Alters Its Anti-Inflammatory Properties

Frontiers in Immunology
Yotam LiorEli C Lewis

Abstract

Human α1-antitrypsin (hAAT) is a 394-amino acid long anti-inflammatory, neutrophil elastase inhibitor, which binds elastase via a sequence-specific molecular protrusion (reactive center loop, RCL; positions 357-366). hAAT formulations that lack protease inhibition were shown to maintain their anti-inflammatory activities, suggesting that some attributes of the molecule may reside in extra-RCL segments. Here, we compare the protease-inhibitory and anti-inflammatory profiles of an extra-RCL mutation (cys232pro) and two intra-RCL mutations (pro357cys, pro357ala), to naïve [wild-type (WT)] recombinant hAAT, in vitro, and in vivo. His-tag recombinant point-mutated hAAT constructs were expressed in HEK-293F cells. Purified proteins were evaluated for elastase inhibition, and their anti-inflammatory activities were assessed using several cell-types: RAW264.7 cells, mouse bone marrow-derived macrophages, and primary peritoneal macrophages. The pharmacokinetics of the recombinant variants and their effect on LPS-induced peritonitis were determined in vivo. Compared to WT and to RCL-mutated hAAT variants, cys232pro exhibited superior anti-inflammatory activities, as well as a longer circulating half-life, despite all three mutated forms ...Continue Reading

References

Jan 1, 1992·Cytotechnology·A CarverA Scott
Jul 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·A L ArchibaldA J Clark
Jun 24, 1988·The American Journal of Medicine·R C Hubbard, R G Crystal
Nov 1, 1987·Journal of Applied Physiology·M A CasolaroG Kawasaki
Jan 1, 1983·British Heart Journal·H GilutzM R Quastel
Jul 29, 1999·American Journal of Respiratory Cell and Molecular Biology·E A EckmanT W Ferkol
Mar 29, 2001·Microbiology and Immunology·Y U KatagiriJ Fujimoto
Aug 15, 2001·Laboratory Investigation; a Journal of Technical Methods and Pathology·A ChurgJ L Wright
Nov 10, 2001·Arteriosclerosis, Thrombosis, and Vascular Biology·S MashibaT Kodama
Feb 26, 2003·American Journal of Respiratory Cell and Molecular Biology·Jérôme Duranton, Joseph G Bieth
Mar 5, 2003·Annual Review of Immunology·Michelle DykstraSusan K Pierce
Apr 15, 2003·Arteriosclerosis, Thrombosis, and Vascular Biology·Philippa J TalmudUNKNOWN Diabetes Atherosclerosis Intervention Study Investigators
Feb 10, 2004·Biochemical and Biophysical Research Communications·Paola Finotti, Andrea Pagetta
Sep 11, 2004·Biochemical and Biophysical Research Communications·Sabina JanciauskieneTim Stevens
Sep 17, 2004·Biochemical and Biophysical Research Communications·Un-Beom KangCheolju Lee
Aug 12, 2005·Proceedings of the National Academy of Sciences of the United States of America·Eli C LewisCharles A Dinarello
Aug 16, 2005·Biochimica Et Biophysica Acta·Koji YoshidaHiroshi Munakata
Jun 15, 2006·Amino Acids·E KarnaukhovaB Golding
Sep 28, 2006·The American Journal of Pathology·Irina PetracheRubin M Tuder
Apr 24, 2007·The International Journal of Biochemistry & Cell Biology·Izabela M NitaSabina M Janciauskiene
Apr 11, 2008·The Journal of Investigative Dermatology·Yuan-Ping HanWarren L Garner
Jun 26, 2009·The New England Journal of Medicine·Edwin K Silverman, Robert A Sandhaus
Feb 24, 2010·Computer Methods and Programs in Biomedicine·Yong ZhangShaofei Xie
Jun 22, 2010·The International Journal of Biochemistry & Cell Biology·Devipriya SubramaniyamSabina Janciauskiene
Nov 10, 2010·The Journal of Clinical Investigation·David A BerginNoel G McElvaney
Jan 25, 2011·Nature Structural & Molecular Biology·Beena Krishnan, Lila M Gierasch
Feb 25, 2011·Journal of Translational Medicine·Christian GrimsteinSihong Song
May 24, 2011·Journal of Molecular and Cellular Cardiology·Stefano ToldoAntonio Abbate
Jun 21, 2011·Methods in Enzymology·Dudley K StricklandToni M Antalis
Dec 29, 2011·Proceedings of the National Academy of Sciences of the United States of America·Isao TawaraPavan Reddy
May 29, 2012·The Journal of Immunology : Official Journal of the American Association of Immunologists·Eyal OzeriEli C Lewis
Jun 16, 2012·Current Molecular Medicine·J M Hunt, R Tuder
Aug 14, 2012·Expert Review of Clinical Immunology·Steven Goldstein, Pavan Reddy
Sep 6, 2012·Proceedings of the National Academy of Sciences of the United States of America·Maria KoulmandaTerry B Strom
Mar 26, 2013·American Journal of Respiratory Cell and Molecular Biology·Angelia D LockettIrina Petrache
Apr 3, 2013·COPD·Christian Mueller, Terence R Flotte

❮ Previous
Next ❯

Methods Mentioned

BETA
PCR
transfection
Protein Assay
flow cytometry
lavage
ELISA
FACS
glycosylation

Clinical Trials Mentioned

NCT01054339
NCT02168686
NCT00377416
NCT00430768

Software Mentioned

FLOWJO
PKsolver
GraphPad Prism

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Proceedings of the National Academy of Sciences of the United States of America
Danny JonigkSabina Janciauskiene
Nihon rinsho. Japanese journal of clinical medicine
T Ishizaki, S Ameshima
© 2022 Meta ULC. All rights reserved