To determine whether insulin regulation of lipolysis is abnormal in subjects with poorly controlled insulin-dependent diabetes mellitus (IDDM), free-fatty acid flux ([1-14C]palmitate) was measured under conditions ranging from complete insulin withdrawal to hyperinsulinemia. Seven nondiabetic and seven IDDM subjects were studied with the pancreatic clamp technique to control plasma insulin, growth hormone, and glucagon concentrations at the desired levels. Preliminary studies were performed to validate the experimental design. The palmitate flux response to insulin withdrawal (2.5 +/- 0.2 vs. 2.5 +/- 0.2 mumol.kg-1.min-1) and maximally antilipolytic insulin concentrations (0.17 +/- 0.02 vs. 0.23 +/- 0.03 mumol.kg-1.min-1) were not different in nondiabetic and IDDM subjects, respectively. In contrast, IDDM subjects required significantly greater plasma free-insulin concentrations to result in equivalent suppression of palmitate flux compared with nondiabetic subjects. Lipolysis was found to be very sensitive to insulin in nondiabetic humans, with half-maximal suppression occurring at plasma free-insulin concentrations of approximately 12 pM (less than 2 microU/ml). We conclude that adipose tissue lipolysis is normally exquisitely sensitive to insulin and that sensitivity, but not responsiveness to insulin, is impaired in poorly controlled IDDM.