The effect of protein intake during gestation and lactation on the lactational performance of primiparous sows was evaluated using 35 Yorkshire x Landrace gilts, allocated to six dietary treatments in a 3 x 2 factorial arrangement. Treatments consisted of three protein levels during gestation, providing approximately 4, 8, and 16 g of lysine/d, and two protein levels (low [L] and high [HI), providing approximately 15 and 45 g of lysine/d, during lactation, respectively. Diets provided similar amounts of ME and all other nutrients. As dietary protein increased during gestation, sows gained more weight and tended to decrease their backfat thickness. There was no gestation x lactation treatment interaction for lactational performance of sows. Feed intake by sows during lactation was usually low but increased (P < .05) with increasing gestation and lactation protein intake and increased linearly (P < .001) as lactation progressed. This linear increase over time was greater (P < .05) in sows fed the H than in sows fed the L protein level. Increased protein intake during lactation reduced (P < .001) 21-d sow weight loss. Milk yield and pig weight gain increased as protein intake increased during gestation (P < .05) and lactation (P < .01). Milk yield did not increase as lactation progressed (P > .05). Pig weight gain increased (P < .05) from wk 1 to 2 of lactation and decreased thereafter. Simple linear regression analysis detected few important relationships between yield of milk components and metabolites or metabolic hormone concentrations. The R2 values for these relationships were < or = .30, except for some relationships between milk component yields and blood urea nitrogen (the range was between .17 and .55). Covariate adjustment for metabolite and metabolic hormone concentrations did not eliminate treatment effects in most cases. This suggests that effects of increased protein intake on milk yield are not fully mediated through metabolite and metabolic hormone concentrations.