Abstract The present study was planned to test the hypothesis that feeding lactating dairy cattle with varying levels of rumen-undegradable protein (RUP) can enhance protein utilization, milk production, milk protein, and nitrogen (N) excretion. Forty mid-lactating crossbred (Jersey × Friesian) cattle were randomly divided into four groups. Four treatment diets were formulated to contain 30%, 40%, 50%, and 60% RUP of crude protein. Dry matter (DM) and crude protein intakes were significantly reduced with increasing dietary RUP levels. Crude protein digestibility increased linearly with incremental increases in dietary RUP levels. Cattle fed 60% RUP showed a linear decrease in N intake compared to that in the other groups. A linear decrease in urinary N and linear increases in net N, milk N, and N-use efficiency were observed with increasing dietary RUP levels. Actual milk, energy-corrected milk, and 4% fat-corrected milk yields (kg/day) increased linearly with an increasing degradability of protein. However, milk protein, solids not fat and total solids, as well as the yields of protein, fat, and lactose, showed significant increases with increased RUP supplementation. Collectively, the results indicate that formulating dairy cow diets to contain 60% RUP results in better lactating performance and N-use efficiency and lower N excretion.