Chapter 13: Crystallization of RNA–Protein Complexes
Eiji Obayashi, Chris Oubridge, Daniel Pomeranz Krummel & Kiyoshi Nagai
RNA-binding proteins play crucial roles in many biological processes, such as transcription, premRNA splicing, nuclear-cytoplasmic transport of RNA, and translation of mRNA. Specific RNA–protein interactions are key to the correct assembly of ribonucleoprotein complexes and their biological functions. To date, more than 100 unique RNA–protein crystals have been prepared and there are more than 300 entries of RNA–protein complex structures in the Protein Data Bank. This chapter focuses on methods of RNA–protein complex crystallization discussed in six sections: determination of protein-binding sites in RNA, preparation of RNA, preparation of protein, annealing of RNA, reconstitution of RNA–protein complex, and searching crystallization conditions.