Low affinity integrin states have faster ligand binding kinetics than the high affinity state

  1. Jing Li
  2. Jiabin Yan
  3. Timothy A Springer  Is a corresponding author
  1. Boston Children's Hospital, United States

Abstract

Integrin conformational ensembles contain two low-affinity states, bent-closed and extended-closed, and an active, high-affinity, extended-open state. It is widely thought that integrins must be activated before they bind ligand; however, one model holds that activation follows ligand binding. As ligand-binding kinetics are not only rate limiting for cell adhesion but also have important implications for the mechanism of activation, we measure them here for integrins α4β1 and α5β1 and show that the low-affinity states bind substantially faster than the high-affinity state. On and off-rates are similar for integrins on cell surfaces and as ectodomain fragments. Although the extended-open conformation's on-rate is ~20-fold slower, its off-rate is ~25,000-fold slower, resulting in a large affinity increase. The tighter ligand-binding pocket in the open state may slow its on-rate. Low affinity integrin states not only bind ligand more rapidly, but are also more populous on the cell surface than high affinity states. Thus, our results suggest that integrin binding to ligand may precede, rather than follow, activation by 'inside-out signaling'.

Data availability

All data generated or analyzed during this study are included in the manuscript and source data files submitted.

Article and author information

Author details

  1. Jing Li

    Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Jiabin Yan

    Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Timothy A Springer

    Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, United States
    For correspondence
    springer@crystal.harvard.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6627-2904

Funding

National Heart, Lung, and Blood Institute (R01-HL131729)

  • Jing Li
  • Timothy A Springer

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Reinhard Fässler, Max Planck Institute of Biochemistry, Germany

Publication history

  1. Preprint posted: July 26, 2021 (view preprint)
  2. Received: August 25, 2021
  3. Accepted: December 1, 2021
  4. Accepted Manuscript published: December 2, 2021 (version 1)
  5. Version of Record published: January 5, 2022 (version 2)

Copyright

© 2021, Li et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Jing Li
  2. Jiabin Yan
  3. Timothy A Springer
(2021)
Low affinity integrin states have faster ligand binding kinetics than the high affinity state
eLife 10:e73359.
https://doi.org/10.7554/eLife.73359
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