Abstract: In this study, the aim was to determine the influence of ultrasound-assisted phosphorylation on the physicochemical properties of chickpea protein isolates (CPI). Three aspects of the effects of combined treatment on the properties of CPI were assessed: physicochemical properties, molecular structure, and functional characteristics. The results showed that with the increase of ultrasound intensity, the physicochemical properties and structure of CPI change significantly. Among them, ultrasound-assisted phosphorylation at 300 W led to a maximum phosphate content of 10.63 μg/mL in the proteins, while increasing the solubility to 89.02% and yielding turbidity of 1.98. In addition, both sulfhydryl contents and surface hydrophobicity were decreased to 10.48 μmol/g and 80.5, accompanied by changes in the secondary structure of the proteins. These findings indicate that ultrasound promotes effective phosphorylation. Hydrophobic aggregation caused by protein unfolding was mitigated by electrostatic stabilization, thereby enhancing the emulsification and foaming properties of CPI. Multiscale analyses demonstrated that ultrasound-assisted phosphorylation effectively enhances the physicochemical properties and structural characteristics of CPI molecules. The findings of this study provide a theoretical basis for advancing food protein modification technologies.