Fibroblast growth factor 21 (FGF21) is a novel hormone-like polypeptide that when administered exogenously, has been shown to have beneficial effects on food intake, body weight, and metabolism. The in vivo mechanisms of action for its positive metabolic effects remain to be fully elucidated. It has been shown that PEGylation of human FGF21 at specific and preferred sites confer superior metabolic pharmacology. We therefore hypothesized that low doses of PEGylated (30 K PEG on position Q108) FGF21 (PEG30-Q108) would improve insulin action, independent of any effect on food intake or body weight. We identified a dose (0.25 mg/kg) that had no effect on food intake or body weight, yet did show beneficial metabolic effects. Four groups of 12 weeks, high-fat fed, insulin resistant mice were studied: mice dosed subcutaneously once with vehicle or 0.25 mg/kg of PEG30-Q108 24 h before the experiment, or mice dosed 4 times over 2 weeks with vehicle or PEG30-Q108. Conscious, unrestrained mice were fasted for 5 h and underwent a hyperinsulinemic–euglycemic clamp. Both PEG30-Q108 treatments significantly lowered fasting insulin compared to vehicle, with no difference in food intake or body weight. Insulin-stimulated whole body glucose utilization was normalized to that of lean mice with both PEG30-Q108 treatments compared to vehicle. This accounted for all of the enhanced insulin action, as there was no improvement in insulin's ability to suppress endogenous glucose production. In line with these findings, neither PEG30-Q108 treatment lowered hepatic triglycerides. These results demonstrate the profound ability of PEG30-Q108 to increase whole body insulin sensitivity.