Lessons learned from Sample Preparation and Structural Analysis of Three Membrane Channels

Sample preparation and data analysis are important steps in achieving high-resolution cryo-EM structures. In this talk, I will discuss the lessons and tips that I learned while studying three different membrane channels, including the proton-activated chloride channel (PAC), pannexin 1 (PANX1) and transient receptor potential cation channel subfamily M member 5 (TRPM5). I will cover topics including detergent/nanodisc optimization, the impact of GFP tag on sample quality, and data processing strategies to overcome various issues such as featureless small target, conformational heterogeneity, and protein flexibility. Finally, I will also provide some tips that we often use to boost map quality and resolution. I hope my experiences will be of value to your projects and help you improve the reconstruction of cryo-EM structures.