Identifying molecular mechanisms regulating cellular energy utilization may lead to increased understanding of maintenance energy cost and improved feed efficiency in beef cows. Three experiments were conducted to characterize measures of residual feed intake (RFI) in pregnant beef cows; to examine the effects of moderate dietary restriction on visceral organ mass and proteins relating to energy metabolism; and to investigate the influence of pregnancy on visceral organ mass and proteins relating to energy metabolism. The first experiment combined data from five experiments using 321 pregnant Angus × Simmental cows. Including ultrasound fat measures and diet/management information increased the feed intake prediction model R2 by 7.3% and > 20%, respectively. Individual experiment RFI models varied greatly in accuracy. In the second experiment, 22 pregnant beef cows were fed at 85% (LOW; n=11) or 140% (HIGH; n=11) of net energy requirements during mid- to late-gestation. Tissue samples from liver, kidney, muscle, ruminal papillae, pancreas, and small intestinal muscosa were collected. Western blots were conducted to quantify abundance of: proliferating cell nuclear antigen, ATP synthase, ubiquitin, and Na/K+ ATPase for all tissues; peroxisome proliferator-activated receptor gamma, peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α), and 5’-adenosine monophosphate-activated protein kinase and phosphorylated-AMPK (pAMPK) for liver, muscle, and rumen; phosphoenolpyruvate carboxykinase for liver and kidney; and uncoupling protein 2 for liver. Cows fed HIGH had greater (P ≤ 0.04) ADG and final BW than cows fed LOW. Ubiquitin abundance in muscle was greater (P = 0.009) in cows fed LOW, and PCG-1α in liver was greater (P = 0.03) in cows fed HIGH. In the third experiment, 18 pregnant (PREG; n =9) or non-pregnant (OPEN; n=9) Angus × Simmental cows were fed for ad libitum intake during mid- to late-gestation. Tissues were weighed and collected and analyzed for protein abundance as described in the second experiment. Liver mass was lower (P ≤ 0.02), abundance of Na+/K+-ATPase was greater (P =0.04) and rumen pAMPK abundance was increased (P = 0.006) in PREG cows. These experiments indicate that measuring RFI in pregnant cows may pose some challenges, and nutrient restriction and pregnancy can influence molecular factors influencing feed efficiency. / Financial support was provided by Canadian Beef Cattle Industry Science Cluster, through funding provided by the Beef Cattle Research Council and Agriculture and Agri-Food Canada, Agriculture Adaptation Council-Farm Innovation Program, Ontario Ministry of Agriculture, Food and Rural Affairs, and the Ontario Cattleman’s Association.