Aim. The results of numerous current studies indicate that vitamin B₁ (thiamine, Th) is a satellite of acetylcholine (ACh) in nerve cells and neuromuscular junctions. However, the mechanisms underlying the relationship between the functions of these compounds have not been fully elucidated. One of the modern approaches to resolve successfully this issue is the study of target proteins of both ligands using the combination of complex traditional biochemical methods and technologies of structural biology. Conclusions. These results broaden the understanding of the molecular mechanisms of vitamin B₁ functioning. It’s important for the development of treatment strategies for neurodegenerative and other diseases. Bioinformatics tools made it possible to predict the significance of the interaction of Th with proteins of the Agrin–LRP4 complex (ThBP): 1) stabilization of the Agrin–LRP4 complex; 2) implementation of the functions of the mobile Th pool [1] by the nAChR cluster; 3) hydrolysis of Th phosphates by proteins of the nAChR cluster.