Luminescent sensing of conformational integrin activation in living cells. Cell Rep 2025 Feb 25;44(2):115319
Date
02/18/2025Pubmed ID
39964812Pubmed Central ID
PMC11861568DOI
10.1016/j.celrep.2025.115319Scopus ID
2-s2.0-85217773499 (requires institutional sign-in at Scopus site) 2 CitationsAbstract
Integrins are major receptors for secreted extracellular matrix, playing crucial roles in physiological and pathological contexts, such as angiogenesis and cancer. Regulation of the transition between inactive and active conformation is key for integrins to fulfill their functions, and pharmacological control of those dynamics may have therapeutic applications. We create and validate a prototypic luminescent β1 integrin activation sensor (β1IAS) by introducing a split luciferase into an activation reporting site between the βI and the hybrid domains. As a recombinant protein in both solution and living cells, β1IAS accurately reports β1 integrin activation in response to (bio)chemical and physical stimuli. A short interfering RNA (siRNA) high-throughput screening on live β1IAS knockin endothelial cells unveils hitherto unknown regulators of β1 integrin activation, such as β1 integrin inhibitors E3 ligase Pja2 and vascular endothelial growth factor B (VEGF-B). This split-luciferase-based strategy provides an in situ label-free measurement of integrin activation and may be applicable to other β integrins and receptors.
Author List
Villari G, Gioelli N, Gino M, Zhang H, Hodge K, Cordero F, Zanivan S, Zhu J, Serini GAuthor
Jieqing Zhu PhD Professor in the Biochemistry department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Human Umbilical Vein Endothelial CellsHumans
Integrin beta1
Luciferases
Luminescent Measurements
Protein Conformation
RNA, Small Interfering
