Titin-Based Nanoparticle Tension Sensors Map High-Magnitude Integrin Forces within Focal Adhesions

• Type of Material: Article
• Authors: Kornelia Galior, Emory University; Yang Liu, Emory University; Kevin Yehl, Emory University; Skanda Vivek, Emory University; Khalid Salaita, Emory University
• Language: English
• Date: 2016-01-01
• Publisher: American Chemical Society
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2015 American Chemical Society
• Final Published Version (URL): http://dx.doi.org/10.1021/acs.nanolett.5b03888
• Title of Journal or Parent Work: Nano Letters
• ISSN: 1530-6984
• Volume: 16
• Issue: 1
• Start Page: 341
• End Page: 348
• Grant/Funding Information: K.S. acknowledges support from the NIH (R01-GM097399), the Alfred P. Sloan Research Fellowship, and the NSF CAREER award.
• Keywords: PROTEIN; Science & Technology; Integrins; Materials Science; Science & Technology - Other Topics; Chemistry; Physics; Physics, Applied; MECHANOTRANSDUCTION; Physics, Condensed Matter; Nanoscience & Nanotechnology; MIGRATION; CELL-ADHESION; Chemistry, Physical; Technology; MECHANICAL TENSION; DYNAMICS; mechanotransduction; PROBES; biophysics; ARCHITECTURE; IMMUNOGLOBULIN DOMAINS; Materials Science, Multidisciplinary; MICROSCOPY; focal adhesion; Physical Sciences; Chemistry, Multidisciplinary
• Research Categories: Chemistry, General; Physics, General

Abstract

Mechanical forces transmitted through integrin transmembrane receptors play important roles in a variety of cellular processes ranging from cell development to tumorigenesis. Despite the importance of mechanics in integrin function, the magnitude of integrin forces within adhesions remains unclear. Literature suggests a range from 1 to 50 pN, but the upper limit of integrin forces remains unknown. Herein we challenge integrins with the most mechanically stable molecular tension probe, which is comprised of the immunoglobulin 27th (I27) domain of cardiac titin flanked with a fluorophore and gold nanoparticle. Cell experiments show that integrin forces unfold the I27 domain, suggesting that integrin forces exceed 30-40 pN. The addition of a disulfide bridge within I27 clamps the probe and resists mechanical unfolding. Importantly, incubation with a reducing agent initiates SH exchange, thus unclamping I27 at a rate that is dependent on the applied force. By recording the rate of S-S reduction in clamped I27, we infer that integrins apply 110 ± 9 pN within focal adhesions of rat embryonic fibroblasts. The rates of S-S exchange are heterogeneous and integrin subtype-dependent. Nanoparticle titin tension sensors along with kinetic analysis of unfolding demonstrate that a subset of integrins apply tension many fold greater than previously reported.