In this study, myofibrillar proteins (MPs) of thawed pork were treated with plasma-activated water (PAW) for diverse durations (0, 5, 10, 15, 20, and 25 s) to investigate whether the function of MPs improved by PAW and the corresponding regulatory mechanism. The results found that PAW treatments increased the surface hydrophobicity, altered secondary and tertiary structure of MPs. The α-helix content of MPs treated by PAW reduced from 37.3% to 31.25%. In PAW25s group, the oxidation of MPs significantly raised reflected by the higher carbonyl content and lower total sulfhydryl content compared to other groups (P<0.05). Furthermore, PAW treatments increased the whiteness, improved the strength, immobilized water contents, resilience, chewiness, and adhesiveness of MP gels. The observation of intermolecular forces and microstructure of MP gels presented the increase in ionic bonding, disulfide bonding, and hydrophobic interactions, but decrease in hydrogen bonding in MP gels with PAW treatments, leading to more homogeneous and denser gel structures compared to control group. In conclusion, PAW treatment for a short duration significantly fixed and enhanced the function of MPs extracted from thawed pork, and to some extent, improved the processing quality of MPs of thawed pork.