Cryogenic electron microscopy (cryo-EM) has recently emerged as an optimal technique for the determination of histone methyltransferase-nucleosome complex structures. Histone methyltransferases are a group of enzymes that posttranslationally methylate histone lysine and arginine residues on the nucleosome, providing important epigenetic signals that regulate gene expression. Here we describe a protocol to solve the structure of histone lysine methyltransferase Dot1L bound to a chemically ubiquitylated nucleosome, including complex reconstitution, crosslinking, grid preparation, and data collection and analysis. Throughout, we discuss key steps requiring optimization to allow this protocol to serve as a starting point for the determination of new histone methyltransferase-nucleosome complex structures.
CITATION STYLE
Spangler, C. J., & McGinty, R. K. (2022). Determination of Histone Methyltransferase Structures in Complex with the Nucleosome by Cryogenic Electron Microscopy. In Methods in Molecular Biology (Vol. 2529, pp. 149–168). Humana Press Inc. https://doi.org/10.1007/978-1-0716-2481-4_8
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