Voltage-gated sodium channels (Navs) are key players in neuronal excitability and pain signaling. Precise gating of these channels is crucial as even small functional alterations can lead to pathological phenotypes such as pain or heart failure. Mechanical stress has been shown to affect sodium channel gating. This suggests that components which stabilize the channel are necessary to ensure precise channel gating in living organisms. We used whole cell patch clamp of Nav1.7 heterologously expressed in HEK cells in presence and absence of the β1 subunit to study the impact of the β1 subunit on mechanical susceptibility of Nav1.7. All groups were measured in the presence and absence of mechanical stress applied by the flow of extracellular solution on the patched cell via a gravity-driven perfusion system. We show that mechanical shear stress shifts voltage dependence of activation (ΔVhalf= 10.5 mV, p= 0.0004), and of fast inactivation ((ΔVhalf= 6.3 mV, p=0.04) to more depolarized potentials. Co-expression of the β1 subunit, however, protects both voltage dependence of activation and fast inactivation of Nav1.7 against mechanical shear stress. To study the underlying mechanisms enabling the β1 subunit to mechanically stabilize Nav1.7, we used molecular dynamics simulations (MD), homology modelling and site-directed mutagenesis. MD simulations were carried out using GROMACS 2016 using the β1 structure published by Yan et al. 2017. Simulations of WT β1 and the C43A mutant revealed an intramolecular disulfide bond (Cys21-Cys43) to be crucial for tight binding of the subunit to the channel. Using a homology model of Nav1.7 based on the Nav1.4β1 Complex (Yan et al. 2017), we show that β1 binds to segment 6 of domain IV of Nav1.7. . To investigate the functional impact of the mutation we generated a C43A β1 subunit by site-directed mutagenesis. Whole cell patch clamp experiments with the C43A β1 subunit and Nav1.7 show that the mutant prevents mechanical modulation of voltage dependence of activation, but not of fast inactivation. Our data reveal a novel feature of β subunit-mediated ion channel modfication: the β1 subunit is able to influence the mechanical susceptibility of Nav1.7. Furthermore our patch data emphasize the unique role of segment 6 of domain IV for sodium channel fast inactivation and confirm previous reports that fast inactivation can be modified by interfering with the extracellular end of segment 6 of domain IV. Our study suggests that physiological gating of Nav1.7 may be protected against mechanical stress in a living organism by assembly with the β1 subunit.