This study examined the effect of different Prunus mume polyphenol (PMP) doses on the structure and gel properties of bovine myofibrillar proteins in oxidative conditions. The underlying regulatory mechanisms were elucidated using data-independent acquisition quantitative proteomics. The results showed that low-to-moderate PMP concentrations (0.1%, 0.2%, and 0.5%) effectively inhibited both the increase in oxidation-induced carbonyl groups and the decline in sulfhydryl content. These concentrations also prevented a decrease in surface hydrophobicity and particle size, while increasing the absolute zeta potential value. Contrarily, high-dose PMP (1%) reduced the sulfhydryl group levels and absolute zeta potential value, while increasing the surface hydrophobicity and particle aggregation. DIA proteomic analysis further demonstrated that PMP significantly upregulated the structural and functional proteins, including myosin and heat shock proteins, as well as the key enzymes involved in glycolysis. These changes alleviated oxidative damage by enhancing protein synthesis, regulating energy metabolism, and facilitating iron ion transport.