Abstract: This study aims to study the interaction between starch and wheat protein that affects the texture, stability, and sensory properties of flour products. At present, interaction studies mainly focus on the dough system, but there are few reports on the changes of fine structures in solution system. In this study, the interaction mechanism between albumin, globulin, gliadin, gluten and amylose in solution system was studied by ultraviolet spectroscopy, fluorescence spectroscopy, and synchronous fluorescent spectroscopy. The results showed that amylose could effectively quench the endogenous fluorescence of albumin, globulin, gliadin and gluten at different temperatures, and the quenching constant decreased with the increase of temperature, indicating that the quenching mechanism of amylose and the four proteins was statically quenching. Albumin and globulin binded to amylose by hydrogen bond and van der Waals force, which was exothermic and enthalpy driven spontaneous process. The binding constant decreased with the increase of temperature, and the binding force was weak. Gliadin and gluten binded to amylose mainly through hydrophobic interactions and hydrogen bonding, which was endothermic, entropy driven spontaneous process, where binding constant increased with temperature and the binding force was strong. Synchronous fluorescence spectroscopy results indicated that when amylose formed complexes with four proteins, albumin, globulin and gliadin binding sites were closer to tyrosine residues and gluten was closer to tryptophan residues. This study will provide a theoretical basis for the design of starch-based food.