Examining the Effect of Kindlin-3 Binding Site Mutation on LFA-1-ICAM-1 Bonds by Force Measuring Optical Tweezers

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McDonald , C , Morrison , V L , McGloin , D & Fagerholm , S C 2022 , ' Examining the Effect of Kindlin-3 Binding Site Mutation on LFA-1-ICAM-1 Bonds by Force Measuring Optical Tweezers ' , Frontiers in Immunology , vol. 12 , 792813 . https://doi.org/10.3389/fimmu.2021.792813

Title: Examining the Effect of Kindlin-3 Binding Site Mutation on LFA-1-ICAM-1 Bonds by Force Measuring Optical Tweezers
Author: McDonald, Craig; Morrison, Vicky L.; McGloin, David; Fagerholm, Susanna Carola
Contributor organization: Molecular and Integrative Biosciences Research Programme
Faculty Common Matters
Integrins in immunity
Date: 2022-01-26
Language: eng
Number of pages: 7
Belongs to series: Frontiers in Immunology
ISSN: 1664-3224
DOI: https://doi.org/10.3389/fimmu.2021.792813
URI: http://hdl.handle.net/10138/341880
Abstract: Integrins in effector T cells are crucial for cell adhesion and play a central role in cell-mediated immunity. Leukocyte adhesion deficiency (LAD) type III, a genetic condition that can cause death in early childhood, highlights the importance of integrin/kindlin interactions for immune system function. A TTT/AAA mutation in the cytoplasmic domain of the beta 2 integrin significantly reduces kindlin-3 binding to the beta 2 tail, abolishes leukocyte adhesion to intercellular adhesion molecule 1 (ICAM-1), and decreases T cell trafficking in vivo. However, how kindlin-3 affects integrin function in T cells remains incompletely understood. We present an examination of LFA-1/ICAM-1 bonds in both wild-type effector T cells and those with a kindlin-3 binding site mutation. Adhesion assays show that effector T cells carrying the kindlin-3 binding site mutation display significantly reduced adhesion to the integrin ligand ICAM-1. Using optical trapping, combined with back focal plane interferometry, we measured a bond rupture force of 17.85 +/- 0.63 pN at a force loading rate of 30.21 +/- 4.35 pN/s, for single integrins expressed on wild-type cells. Interestingly, a significant drop in rupture force of bonds was found for TTT/AAA-mutant cells, with a measured rupture force of 10.08 +/- 0.88pN at the same pulling rate. Therefore, kindlin-3 binding to the cytoplasmic tail of the beta 2-tail directly affects catch bond formation and bond strength of integrin-ligand bonds. As a consequence of this reduced binding, CD8+ T cell activation in vitro is also significantly reduced.
Subject: LFA-1
kindlin-3
T cell
bond strength
ICAM-1
FUNCTION-ASSOCIATED ANTIGEN-1
T-CELL-ACTIVATION
INTEGRIN
LIGAND
ADHESION
1182 Biochemistry, cell and molecular biology
3121 General medicine, internal medicine and other clinical medicine
Peer reviewed: Yes
Rights: cc_by
Usage restriction: openAccess
Self-archived version: publishedVersion


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