Millet glutenin and gliadin was obtained by alkali extraction and acid precipitation and millet gliadin was distracted by ethanol. Then their structural characteristics were analyzed. SDS-PAGE electrophoresis results showed that the molecular weight of the millet gliadin was small, while millet glutenin was distributed from high molecular weight to low molecular weight. Furthermore, the XRD analysis showed that both the two kinds of proteins were amorphous structures. The secondary structure was measured by circular dichroism. Millet gliadin belonged to all alpha structure, and its order was better than that of millet alkali soluble protein. Moreover, by atomic force microscopy analysis, millet gliadin was spherical and its distribution was rather dispersed. In contrast, the distribution of millet glutenin was more concentrated.