Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron MicroscopyEmery Haley *1, Wooyoung Choi *1, Chen Fan 1, Weinan Sun 2,3, Juan Du 1, Wei Lü 1
1Van Andel Institute, 2Vollum Institute, 3Janelia Research Campus
This protocol describes techniques used to determine ion channel structures by cryo-electron microscopy, including a baculovirus system used to efficiently express genes in mammalian cells with minimum effort and toxicity, protein extraction, purification, and quality checking, sample grid preparation and screening, as well as data collection and processing.
