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Differences in self-association between kindlin-2 and kindlin-3 are associated with differential integrin binding
Yasmin A Kadry1, Eesha M Maisuria2, Clotilde Huet-Calderwood1
1Department of Pharmacology, Yale University, New Haven, Connecticut, USA.
The Journal of Biological Chemistry
|June 18, 2020
Summary
Kindlin proteins, crucial for cell adhesion, can self-associate. This oligomerization inhibits their ability to bind integrins and localize to focal adhesions, revealing a novel regulatory mechanism.
Area of Science:
- Cell biology
- Molecular and structural biology
- Biochemistry
Background:
- Integrins are transmembrane receptors controlling cell-extracellular matrix interactions.
- Kindlins are cytoplasmic proteins essential for integrin function and signaling.
- Regulation of kindlin scaffolding function remains poorly understood.
Purpose of the Study:
- To investigate the molecular regulation of kindlin scaffolding functions.
- To determine if kindlins self-associate and how this impacts integrin binding.
- To identify regions and mutations affecting kindlin self-association and function.
Main Methods:
- GFP nanotrap association assays
- Pulldown and integrin-binding assays
- Live-cell imaging
- Chimeric mapping and comparative sequence analysis
Main Results:
- Full-length kindlins (kindlin-2 and kindlin-3) oligomerize in mammalian cells.
- Kindlin self-association inhibits integrin binding and focal adhesion localization.
- F2PH and F3 subdomains are critical for kindlin self-association.
- Specific kindlin-3 mutations reduce self-association, enhance integrin binding, and promote focal adhesion localization.
Conclusions:
- Kindlin self-association acts as a negative regulator of integrin binding.
- Oligomerization state influences kindlin localization and function at focal adhesions.
- Understanding kindlin self-association provides insights into integrin-mediated cell adhesion regulation.
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