Allosteric β1 Integrin Antibodies That Stabilize the Low Affinity State by Preventing the Swing-out of the Hybrid Domain
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The ligand binding function of integrins can be modulated by various monoclonal antibodies by both direct and indirect mechanisms. We have characterized an anti-beta(1) antibody, SG/19, that had been reported to inhibit the function of the beta(1) integrin on the cell surface. SG/19 recognized the wild type beta(1) subunit that exists in a conformational equilibrium between the high and low affinity states but bound poorly to a mutant beta(1) integrin that had been locked in a high affinity state. Epitope mapping of SG/19 revealed that Thr(82) in the beta(1) subunit, located at the outer face of the boundary between the I-like and hybrid domains, was the key binding determinant for this antibody. Direct visualization of the alpha (5)beta(1) headpiece fragment in complex with SG/19 Fab with electron microscopy confirmed the location of the binding surface and showed that the ligand binding site is not occluded by the bound Fab. Surface plasmon resonance showed that alpha (5)beta(1) integrin bound by SG/19 maintained a low affinity toward its physiological ligand fibronectin (Fn) whereas binding by function-blocking anti-alpha(5) antibodies resulted in a complete loss of fibronectin binding. Thus a class of the anti-beta antibodies represented by SG/19 attenuate the ligand binding function by restricting the conformational shift to the high affinity state involving the swing-out of the hybrid domain without directly interfering with ligand docking.
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The SCD of ANGPT2 interacts with β1-integrin, mediated through the hybrid/PSI domain of β1-integrin.
“Interestingly, we found that the SCD of ANGPT2 additionally interacted with β1-integrin. The interaction was mediated through the hybrid/PSI domain of β1-integrin, which is known to undergo conformational changes during the transition of α5β1-integrin from an inactive, half-bent conformation to an active extended conformation (57).”
From Inactive β1-integrin acts as a junctional scaffold for angiopoietin/TIE2/FOXO1 signaling (2026) · cites this paper as [57]
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- Authors
- Luo, B.-H., Strokovich, K., Walz, T., Springer, T. A., & Takagi, J.
- Affiliations
- Harvard University (US)
- Institute for Protein Research, The University of Osaka (JP)
- Corresponding author
- Takagi, Junichi
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2004-06
- Volume
- 279
- Issue
- 26
- Pages
- 27466-27471
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2159165113
- Cited by
- 73
- References
- 38
- Citations by year
- 2026: 3 · 2024: 1 · 2022: 2 · 2021: 3 · 2019: 1 · 2017: 4 · 2016: 2 · 2015: 3 · 2014: 5 · 2013: 5 · 2012: 5
- FWCI
- 3.58
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Civil War Book Review (submittedVersion)
- Funders
- National Heart, Lung, and Blood Institute (HL48675, P01HL048675)
- National Institute of General Medical Sciences (P01GM062580, GM-62580)
- Topics
- Health Sciences › Medicine › Immunology and Allergy › Cell Adhesion Molecules Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Keywords
- beta1 integrin, hybrid domain, high-affinity state, alpha5beta1 integrin, fibronectin binding, ligand binding sites, epitope mapping
- MeSH terms
- Allosteric Regulation, Animals, Antibodies, Monoclonal, Antigen-Antibody Complex, Binding, Competitive, Cricetulus, Cricetinae, Humans, Immunoglobulin G, Models, Molecular, Recombinant Proteins, CHO Cells, Protein Structure, Tertiary, Epitope Mapping, Integrin beta1, Amino Acid Substitution, Surface Plasmon Resonance, Protein Subunits, Mice, Rats
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