Structural basis of human full-length kindlin-3 homotrimer in an auto-inhibited state

• Main Authors: Bu, Wenting, Levitskaya, Zarina, Loh, Zhi Yang, Jin, Shengyang, Basu, Shibom, Ero, Rya, Yan, Xinfu, Wang, Meitian, Ngan, So Fong Cam, Sze, Siu Kwan, Tan, Suet-Mien, Gao, Yong-Gui
• Other Authors: School of Biological Sciences
• Format: Article
• Language: English
• Published: 2021
• Subjects: Science::Biological sciences; Kindlin; FERM
• Online Access: https://hdl.handle.net/10356/146721
• Institution: Nanyang Technological University

Kindlin-1, -2, and -3, directly bind integrin β cytoplasmic tails to regulate integrin activation and signaling. Despite their functional significance and link to several diseases, structural information on full-length kindlin proteins remains unknown. Here, we report the crystal structure of human full-length kindlin-3, which reveals a novel homotrimer state. Unlike kindlin-3 monomer, which is the major population in insect and mammalian cell expression systems, kindlin-3 trimer does not bind integrin β cytoplasmic tail as the integrin-binding pocket in the F3 sub-domain of one protomer is occluded by the PH domain of another protomer, suggesting that kindlin-3 is auto-inhibited upon trimer formation. This is also supported by functional assays in which kindlin-3 knockout K562 erythroleukemia cells re-constituted with the mutant kindlin-3 containing trimer-disrupting mutations exhibited an increase in integrin-mediated adhesion and spreading on fibronectin compared to those re-constituted with wild type kindlin-3. Taken together, our findings reveal a novel mechanism of kindlin auto-inhibition that involves its homotrimer formation.