An allosteric redox switch in domain V of β2-glycoprotein I controls membrane binding and anti-domain I autoantibody recognition

Kumar, Suresh; Chinnaraj, Mathivanan; Planer, William; Zuo, Xiaobing; Macor, Paolo; Tedesco, Francesco; Pozzi, Nicola

doi:10.1016/j.jbc.2021.100890

Title: An allosteric redox switch in domain V of β2-glycoprotein I controls membrane binding and anti-domain I autoantibody recognition

Journal Article · Sun Aug 01 00:00:00 EDT 2021 · Journal of Biological Chemistry

DOI:https://doi.org/10.1016/j.jbc.2021.100890· OSTI ID:1804127

Kumar, Suresh;

;

; Zuo, Xiaobing;

;

β2-glycoprotein I (β₂GPI) is an abundant multidomain plasma protein that plays various roles in the clotting and complement cascades. It is also the main target of antiphospholipid antibodies (aPL) in the acquired coagulopathy known as antiphospholipid syndrome (APS). Previous studies have shown that β₂GPI adopts two interconvertible biochemical conformations, oxidized and reduced, depending on the integrity of the disulfide bonds. However, the precise contribution of the disulfide bonds to β₂GPI structure and function is unknown. Here, we substituted cysteine residues with serine to investigate how the disulfide bonds C32-C60 in domain I (DI) and C288-C326 in domain V (DV) regulate β₂GPI’s structure and function. Results of our biophysical and biochemical studies support the hypothesis that the C32-C60 disulfide bond plays a structural role, whereas the disulfide bond C288-C326 is allosteric. We demonstrate that absence of the C288-C326 bond, unlike absence of the C32-C60 bond, diminishes membrane binding without affecting the thermodynamic stability and overall structure of the protein, which remains elongated in solution. We also document that, while absence of the C32- C60 bond directly impairs recognition of β₂GPI by pathogenic anti-DI antibodies, absence of the C288-C326 disulfide bond is sufficient to abolish complex formation in the presence of anionic phospholipids. We conclude that the disulfide bond C288-C326 operates as a molecular switch capable of regulating β₂GPI’s physiological functions in a redox-dependent manner. We propose that in APS patients with anti-DI antibodies, selective rupture of the C288-C326 disulfide bond may be a valid strategy to lower the pathogenic potential of aPL.

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Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1804127

Alternate ID(s):: OSTI ID: 1815528

Journal Information:: Journal of Biological Chemistry, Journal Name: Journal of Biological Chemistry Vol. 297 Journal Issue: 2; ISSN 0021-9258

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

References (55)

The Pathogenesis of the Antiphospholipid Syndrome Giannakopoulos, Bill; Krilis, Steven A. New England Journal of Medicine, Vol. 368, Issue 11 https://doi.org/10.1056/NEJMra1112830	journal	March 2013
The significance of autoantibodies against β2-glycoprotein I de Groot, Philip G.; Urbanus, Rolf T. Blood, Vol. 120, Issue 2 https://doi.org/10.1182/blood-2012-03-378646	journal	July 2012
IgG antibodies that recognize epitope Gly40-Arg43 in domain I of β2–glycoprotein I cause LAC, and their presence correlates strongly with thrombosis de Laat, Bas; Derksen, Ronald H. W. M.; Urbanus, Rolf T. Blood, Vol. 105, Issue 4 https://doi.org/10.1182/blood-2004-09-3387	journal	February 2005
Chemical synthesis and characterization of wild-type and biotinylated N-terminal domain 1-64 of β2-glycoprotein I Pozzi, Nicola; Banzato, Alessandra; Bettin, Samuele Protein Science, Vol. 19, Issue 5 https://doi.org/10.1002/pro.387	journal	March 2010
Heparin Inhibits the Binding of β2-glycoprotein I to Phospholipids and Promotes the Plasmin-mediated Inactivation of This Blood Protein Guerin, Jan; Sheng, Yonghua; Reddel, Stephen Journal of Biological Chemistry, Vol. 277, Issue 4 https://doi.org/10.1074/jbc.M110176200	journal	January 2002
A non–complement-fixing antibody to β2 glycoprotein I as a novel therapy for antiphospholipid syndrome Agostinis, Chiara; Durigutto, Paolo; Sblattero, Daniele Blood, Vol. 123, Issue 22 https://doi.org/10.1182/blood-2013-11-537704	journal	May 2014
Crystal structure of human β2-glycoprotein I: implications for phospholipid binding and the antiphospholipid syndrome Schwarzenbacher, Robert; Zeth, Kornelius; Diederichs, Kay The EMBO Journal, Vol. 18, Issue 22 https://doi.org/10.1093/emboj/18.22.6228	journal	November 1999
Discovery and characterization of 2 novel subpopulations of aPS/PT antibodies in patients at high risk of thrombosis Chinnaraj, Mathivanan; Planer, William; Pengo, Vittorio Blood Advances, Vol. 3, Issue 11 https://doi.org/10.1182/bloodadvances.2019030932	journal	June 2019
Fibrinogen function achieved through multiple covalent states Butera, Diego; Hogg, Philip J. Nature Communications, Vol. 11, Issue 1 https://doi.org/10.1038/s41467-020-19295-7	journal	October 2020
Adhesion mechanism of human beta 2-glycoprotein I to phospholipids based on its crystal structure Bouma, B. The EMBO Journal, Vol. 18, Issue 19 https://doi.org/10.1093/emboj/18.19.5166	journal	October 1999
Platelet Protein Disulfide Isomerase Promotes Glycoprotein Ibα–Mediated Platelet-Neutrophil Interactions Under Thromboinflammatory Conditions Li, Jing; Kim, Kyungho; Jeong, Si-Yeon Circulation, Vol. 139, Issue 10 https://doi.org/10.1161/CIRCULATIONAHA.118.036323	journal	March 2019
Complement activity and complement regulatory gene mutations are associated with thrombosis in APS and CAPS Chaturvedi, Shruti; Braunstein, Evan M.; Yuan, Xuan Blood, Vol. 135, Issue 4 https://doi.org/10.1182/blood.2019003863	journal	January 2020
Platelets are required for enhanced activation of the endothelium and fibrinogen in a mouse thrombosis model of APS Proulle, Valerie; Furie, Richard A.; Merrill-Skoloff, Glenn Blood, Vol. 124, Issue 4 https://doi.org/10.1182/blood-2014-02-554980	journal	July 2014
β ₂ -Glycoprotein I binds to thrombin and selectively inhibits the enzyme procoagulant functions Pozzi, N.; Acquasaliente, L.; Frasson, R. Journal of Thrombosis and Haemostasis, Vol. 11, Issue 6 https://doi.org/10.1111/jth.12238	journal	June 2013
The J-elongated conformation of β2-glycoprotein I predominates in solution: implications for our understanding of antiphospholipid syndrome Ruben, Eliza; Planer, William; Chinnaraj, Mathivanan Journal of Biological Chemistry, Vol. 295, Issue 31 https://doi.org/10.1074/jbc.RA120.013939	journal	July 2020
The Local Phospholipid Environment Modulates the Activation of Blood Clotting Shaw, Andrew W.; Pureza, Vincent S.; Sligar, Stephen G. Journal of Biological Chemistry, Vol. 282, Issue 9 https://doi.org/10.1074/jbc.M607973200	journal	March 2007
Redox Control of Platelet Aggregation ^† Essex, David W.; Li, Mengru Biochemistry, Vol. 42, Issue 1 https://doi.org/10.1021/bi0205045	journal	January 2003
Measuring and increasing protein stability Pace, C. Trends in Biotechnology, Vol. 8 https://doi.org/10.1016/0167-7799(90)90146-O	journal	January 1990
The autoactivation of factor XII (Hageman factor) induced by low-Mr heparin and dextran sulphate. The effect of the Mr of the activating polyanion Silverberg, M.; Diehl, S. V. Biochemical Journal, Vol. 248, Issue 3 https://doi.org/10.1042/bj2480715	journal	December 1987
Beta 2 glycoprotein I is a substrate of thiol oxidoreductases Passam, Freda H.; Rahgozar, Soheila; Qi, Miao Blood, Vol. 116, Issue 11 https://doi.org/10.1182/blood-2010-02-271494	journal	September 2010
Regulators of complement activity mediate inhibitory mechanisms through a common C3b‐binding mode Forneris, Federico; Wu, Jin; Xue, Xiaoguang The EMBO Journal, Vol. 35, Issue 10 https://doi.org/10.15252/embj.201593673	journal	March 2016
The use of fluorescence methods to monitor unfolding transitions in proteins Eftink, M. R. Biophysical Journal, Vol. 66, Issue 2 https://doi.org/10.1016/S0006-3495(94)80799-4	journal	February 1994
Protein disulfide isomerase inhibition blocks thrombin generation in humans by interfering with platelet factor V activation Stopa, Jack D.; Neuberg, Donna; Puligandla, Maneka JCI Insight, Vol. 2, Issue 1 https://doi.org/10.1172/jci.insight.89373	journal	January 2017
β2-Glycoprotein I can exist in 2 conformations: implications for our understanding of the antiphospholipid syndrome Ağar, Çetin; van Os, Gwendolyn M. A.; Mörgelin, Matthias Blood, Vol. 116, Issue 8 https://doi.org/10.1182/blood-2009-12-260976	journal	August 2010
Specific domain V reduction of beta-2-glycoprotein I induces protein flexibility and alters pathogenic antibody binding Buchholz, Ina; McDonnell, Thomas; Nestler, Peter Scientific Reports, Vol. 11, Issue 1 https://doi.org/10.1038/s41598-021-84021-2	journal	February 2021
How do antiphospholipid antibodies bind ?2-glycoprotein I? Giles, Ian P.; Isenberg, David A.; Latchman, David S. Arthritis & Rheumatism, Vol. 48, Issue 8 https://doi.org/10.1002/art.11101	journal	August 2003
Anticardiolipin antibodies (ACA) directed not to cardiolipin but to a plasma protein cofactor Galli, M.; Barbui, T.; Comfurius, P. The Lancet, Vol. 335, Issue 8705 https://doi.org/10.1016/0140-6736(90)91374-J	journal	June 1990
Immune responses against domain I of β2-glycoprotein I are driven by conformational changes: Domain I of β2-glycoprotein I harbors a cryptic immunogenic epitope de Laat, Bas; van Berkel, Miranda; Urbanus, Rolf T. Arthritis & Rheumatism, Vol. 63, Issue 12 https://doi.org/10.1002/art.30633	journal	November 2011
Free Thiol β2-GPI (β-2-Glycoprotein-I) Provides a Link Between Inflammation and Oxidative Stress in Atherosclerotic Coronary Artery Disease Weaver, James C.; Ullah, Inaam; Qi, Miao Arteriosclerosis, Thrombosis, and Vascular Biology, Vol. 40, Issue 11 https://doi.org/10.1161/ATVBAHA.120.315156	journal	November 2020
The purification of reduced β2-glycoprotein I showed its native activity in vitro Zhou, Saijun; Lu, Ming; Zhao, Jiantong Lipids in Health and Disease, Vol. 16, Issue 1 https://doi.org/10.1186/s12944-017-0555-x	journal	September 2017
β2-Glycoprotein I: evolution, structure and function: β2-Glycoprotein I De Groot, P. G.; Meijers, J. C. M. Journal of Thrombosis and Haemostasis, Vol. 9, Issue 7 https://doi.org/10.1111/j.1538-7836.2011.04327.x	journal	July 2011
Anti- 2 glycoprotein I ( 2GPI) autoantibodies recognize an epitope on the first domain of 2GPI Iverson, G. M.; Victoria, E. J.; Marquis, D. M. Proceedings of the National Academy of Sciences, Vol. 95, Issue 26 https://doi.org/10.1073/pnas.95.26.15542	journal	December 1998
Bioorthogonal Chemistry Enables Single‐Molecule FRET Measurements of Catalytically Active Protein Disulfide Isomerase Chinnaraj, Mathivanan; Barrios, David A.; Frieden, Carl ChemBioChem, Vol. 22, Issue 1 https://doi.org/10.1002/cbic.202000537	journal	September 2020
β2-Glycoprotein I: a novel component of innate immunity Ağar, Çetin; de Groot, Philip G.; Mörgelin, Matthias Blood, Vol. 117, Issue 25 https://doi.org/10.1182/blood-2010-12-325951	journal	June 2011
Antibodies to Domain I of β2Glycoprotein I are in close relation to patients risk categories in Antiphospholipid Syndrome (APS) Banzato, A.; Pozzi, N.; Frasson, R. Thrombosis Research, Vol. 128, Issue 6 https://doi.org/10.1016/j.thromres.2011.04.021	journal	December 2011
Autoregulation of von Willebrand factor function by a disulfide bond switch Butera, Diego; Passam, Freda; Ju, Lining Science Advances, Vol. 4, Issue 2 https://doi.org/10.1126/sciadv.aaq1477	journal	February 2018
beta 2-Glycoprotein I: a plasma inhibitor of the contact activation of the intrinsic blood coagulation pathway Schousboe, I. Blood, Vol. 66, Issue 5 https://doi.org/10.1182/blood.V66.5.1086.1086	journal	November 1985
Complement Activation and Thrombin Generation by MBL Bound to β2-Glycoprotein I Durigutto, Paolo; Macor, Paolo; Pozzi, Nicola The Journal of Immunology, Vol. 205, Issue 5 https://doi.org/10.4049/jimmunol.2000570	journal	August 2020
Oxidation of β2-glycoprotein I associates with IgG antibodies to domain I in patients with antiphospholipid syndrome Raimondo, Maria Gabriella; Pericleous, Charis; Radziszewska, Anna PLOS ONE, Vol. 12, Issue 10 https://doi.org/10.1371/journal.pone.0186513	journal	October 2017
Pathogenesis of antiphospholipid syndrome: understanding the antibodies Meroni, Pier Luigi; Borghi, M. Orietta; Raschi, Elena Nature Reviews Rheumatology, Vol. 7, Issue 6 https://doi.org/10.1038/nrrheum.2011.52	journal	May 2011
Complement C5 but not C3 is expendable for tissue factor activation by cofactor-independent antiphospholipid antibodies Müller-Calleja, Nadine; Ritter, Svenja; Hollerbach, Anne Blood Advances, Vol. 2, Issue 9 https://doi.org/10.1182/bloodadvances.2018017095	journal	May 2018
Effect of Heparin on the Activation of Factor XII and the Contact System in Plasma Pixley, Robin A.; Cassello, Anita; De La Cadena, Raul A. Thrombosis and Haemostasis, Vol. 66, Issue 05 https://doi.org/10.1055/s-0038-1646456	journal	January 1991
Phospholipid-Bound β2-Glycoprotein I Induces the Production of Anti-Phospholipid Antibodies Subang, Rebecca; Levine, Jerrold S.; Janoff, Andrew S. Journal of Autoimmunity, Vol. 15, Issue 1 https://doi.org/10.1006/jaut.2000.0382	journal	August 2000
Naturally occurring free thiols within β2-glycoprotein I in vivo: nitrosylation, redox modification by endothelial cells, and regulation of oxidative stress–induced cell injury Ioannou, Yiannis; Zhang, Jing-Yun; Passam, Freda H. Blood, Vol. 116, Issue 11 https://doi.org/10.1182/blood-2009-04-215335	journal	September 2010
Simultaneous characterization of SNPs and N-glycans from multiple glycosylation sites of intact β-2-glycoprotein-1 (B2GP1) by ESI-qTOF-MS Baerenfaenger, Melissa; Meyer, Bernd Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics, Vol. 1867, Issue 6 https://doi.org/10.1016/j.bbapap.2019.03.007	journal	June 2019
Pathogenic anti-β2-glycoprotein I antibodies recognize domain I of β2-glycoprotein I only after a conformational change de Laat, Bas; Derksen, Ronald H. W. M.; van Lummel, Menno Blood, Vol. 107, Issue 5 https://doi.org/10.1182/blood-2005-05-1943	journal	March 2006
Toward Understanding Tryptophan Fluorescence in Proteins Chen, Yu; Barkley, Mary D. Biochemistry, Vol. 37, Issue 28 https://doi.org/10.1021/bi980274n	journal	June 1998
Solution Structure of Human and Bovine β2-Glycoprotein I Revealed by Small-angle X-ray Scattering Hammel, Michal; Kriechbaum, Manfred; Gries, Anna Journal of Molecular Biology, Vol. 321, Issue 1 https://doi.org/10.1016/S0022-2836(02)00621-6	journal	August 2002
Hydroxychloroquine directly reduces the binding of antiphospholipid antibody–β2-glycoprotein I complexes to phospholipid bilayers Rand, Jacob H.; Wu, Xiao-Xuan; Quinn, Anthony S. Blood, Vol. 112, Issue 5 https://doi.org/10.1182/blood-2008-03-144204	journal	September 2008
Polyphosphate: an ancient molecule that links platelets, coagulation, and inflammation Morrissey, James H.; Choi, Sharon H.; Smith, Stephanie A. Blood, Vol. 119, Issue 25 https://doi.org/10.1182/blood-2012-03-306605	journal	June 2012
Novel assays of thrombogenic pathogenicity in the antiphospholipid syndrome based on the detection of molecular oxidative modification of the major autoantigen β ₂ ‐glycoprotein I Ioannou, Yiannis; Zhang, Jing‐Yun; Qi, Miao Arthritis & Rheumatism, Vol. 63, Issue 9 https://doi.org/10.1002/art.30383	journal	August 2011
Redox control of β ₂ -glycoprotein I-von Willebrand factor interaction by thioredoxin-1: Reduced β ₂ GPI and von Willebrand factor Passam, F. H.; Rahgozar, S.; Qi, M. Journal of Thrombosis and Haemostasis, Vol. 8, Issue 8 https://doi.org/10.1111/j.1538-7836.2010.03944.x	journal	August 2010
β2-Glycoprotein I inhibits von Willebrand factor–dependent platelet adhesion and aggregation Hulstein, Janine J. J.; Lenting, Peter J.; de Laat, Bas Blood, Vol. 110, Issue 5 https://doi.org/10.1182/blood-2006-10-053199	journal	September 2007
In vivo distribution of β2 glycoprotein I under various pathophysiologic conditions Agostinis, Chiara; Biffi, Stefania; Garrovo, Chiara Blood, Vol. 118, Issue 15 https://doi.org/10.1182/blood-2011-01-333617	journal	October 2011
Molecular pathophysiology of the antiphospholipid syndrome: the role of oxidative post-translational modification of beta 2 glycoprotein I: Post-translational modification of beta 2 glycoprotein I Passam, F. H.; Giannakopoulos, B.; Mirarabshahi, P. Journal of Thrombosis and Haemostasis, Vol. 9 https://doi.org/10.1111/j.1538-7836.2011.04301.x	journal	July 2011

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Title: An allosteric redox switch in domain V of β2-glycoprotein I controls membrane binding and anti-domain I autoantibody recognition

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