Determination of single-molecule loading rate during mechanotransduction in cell adhesion

Cells connect with their environment through surface receptors and use physical tension in receptor–ligand bonds for various cellular processes. Single-molecule techniques have revealed bond strength by measuring “rupture force,” but it has long been recognized that rupture force is dependent on loading rate—how quickly force is ramped up. Thus, the physiological loading rate needs to be measured to reveal the mechanical strength of individual bonds in their functional context. We have developed an overstretching tension sensor (OTS) to allow more accurate force measurement in physiological conditions with single-molecule detection sensitivity even in mechanically active regions. We used serially connected OTSs to show that the integrin loading rate ranged from 0.5 to 4 piconewtons per second and was about three times higher in leukocytes than in epithelial cells.