We determined whether the amount of protein in the diet can affect insulin requirements in subjects with diabetes mellitus and glucose metabolism in normal subjects. Seven normal-weight volunteers with uncomplicated, intensively controlled, type I (insulin-dependent) diabetes and 12 similar nondiabetic subjects were studied on a metabolic ward before and after consuming a maintenance-energy but protein-free diet for 10 days. Blood glucose levels of diabetic subjects were measured seven times daily in response to insulin administration by continuous subcutaneous infusion. The plasma glucose appearance rate (Ra) was measured in seven normal subjects and all diabetic subjects using a primed-continuous infusion of D-[6,6-2H2]glucose. After adaptation to the protein-restricted diet, diabetic subjects experienced a 30% decrease in average preprandial and average daily blood glucose concentrations (P < .01); this occurred despite a concurrent 25% decrease in both basal and bolus insulin dosages (P < .001). Protein restriction decreased the postabsorptive glucose Ra (P < .05) and insulin concentrations (P < .01) of normal subjects by 20%, and increased their fasting glucagon concentrations by 24% (P < .01). We conclude that severe protein restriction decreases insulin requirements in type I diabetes and fasting hepatic glucose output and basal insulin levels in normal subjects. This effect appears to be mediated in part by decreased hepatic gluconeogenesis, but a contributory influence of increased insulin sensitivity is not ruled out.