Ui Okada, Satoshi Murakami
Three-dimensional structural information is crucial for the precise and detailed investigation of protein functions and their molecular mechanisms. X-ray crystallography is an effective method for determining protein structures. Here, we describe our standardized protocols, including overexpression, purification, crystallization, and handling of crystals for the structural analysis of multidrug transporters. We have determined several crystal structures of resistance-nodulation-division (RND) and ATP-binding cassette (ABC)-type tripartite multidrug transporters using these methods. These results facilitate the understanding of the multidrug resistance molecular mechanisms, which are mediated by tripartite multidrug efflux complexes.