Muscarinic acetylcholine receptor belongs to the G-coupled receptor family, the cooperation between its five subtypes is crucially important in maintaining the normal physiological function. At present, many types of biological resources are increasing, which provides unprecedented opportunities to study the relationships between muscarinic acetylcholine receptor subtypes. In this study, we take advantage of different types of data, using bioinformatics tools and strategies, to analyze the relationship between muscarinic acetylcholine receptor subtypes from four aspects, including evolution, sequence similarity, expression correlation and protein-protein interaction network. From evolution and sequence similarity aspects, we found the five subtypes of muscarinic acetylcholine receptor can be classified into two subclasses. The first subclass includes M1, M3, and M5, and the second subclass includes M2 and M4. The evolutionary distance between two subtypes of the same subclass is relatively near, and the sequence similarity between them is relatively high. From expression correlation aspect, we found that the subtypes of the first subclass have a positive correlation with the subtypes of the second subclass, that is, there are potential co-expression relationships between them. From protein-protein interaction aspect, we found that the subtypes of the first subclass have indirect interactions with the subtypes of the second subclass, which indicates cooperation relationships.