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

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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'.

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

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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.
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.
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.

"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.

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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.