This work investigated the impact of high intensity ultrasound (HIUS) pretreatment on the functional properties and structural characteristics of micellar casein concentrate (MCC). Microfiltered casein protein retentates were treated with HIUS for 0.5, 1, 2, and 5 min prior to spray drying. The results showed that conductivity, solubility, emulsifying, gelling increased significantly as the ultrasonic time prolonged, but the change of pH value were insignificant. In addition, the structural characteristics of MCC for all samples were studied. There was an increase in surface hydrophobicity (Ho) and a reduction in particle size compared with the control (without HIUS pretreatment). The secondary structure of HIUS pretreated MCC samples changed significantly with an increase in β-sheets and random coils and a reduction in α-helix and β-turn. It can be speculated that HIUS pretreatment facilitate the change of functional properties of MCC and these changes would promote its application in food industry.