Talin is an integrin-binding protein located at focal adhesion site which serves as both an adapter and a force transmitter.Its integrin binding activity is regulated by the intermolecular interaction between its F3 and RS domains.Here, we first used atomic force microscope (AFM)to measure the strength of talin autoinhibition complex.Our results show that the lifetime of talin autoinhibition complex does not change significantly at smaller forces, while it drops rapidly at larger force (＞ 10pN).Molecular dynamics (MD) simulations revealed an additional stable conformation of the talin autoinhibition complex other than that observed by X-ray crystallography.Further analysis indicated that forces regulated the equilibrium of the two stable binding states and resulted in the non-exponential force dependence of the binding lifetime.Our findings shed light on a better understanding about the stability of talin autoinhibition and provide a new point of view on the function of talin in focal adhesion.