Insulin may be a potential target of ONOO- during conditions involving accelerated rates of oxygen radical and nitric oxide generation in pancreatic islet beta-cells. To evaluate the effect of protein tyrosine nitration on insulin function, the porcine insulin was nitrated by ONOO-. Insulin nitration was confirmed by spectral changes, Native-PAGE, and mass spectrometry. The bioassay results show that mono-nitro-insulin affected its receptor binding and hypoglycemic capacities to a certain extent, but still hold high biological activities. (c) 2005 Elsevier Inc. All rights reserved.