Poster Presentation The 46th Lorne Conference on Protein Structure and Function 2021

How to routinely get sub 2.5 Angstrom resolution GPCRs structures by single particle cryoEM (#121)

Matthew Belousoff 1 , Patrick Sexton 1 , Denise Wootten 1 , Rado Danev 2
  1. Drug Development Biology, Monash Institute of Pharmaceutical Sciences, Parkville, VIC, Australia
  2. Graduate School of Medicine, Univeristy of Tokyo, Tokyo, Bunkyo-ku, Japan

In the last few years we have been able to see steady improvements in both the sample preparation and data collection strategies for single particle cryoEM analysis of G-protein coupled receptors (GPCRs). What began with structure determinations at medium resolution using the Volta Phase Plate (VPP) have now moved to conventional transmission electron microscope (CTEM) approaches which have enabled us to break the 2 angtrom resolution barrier and have allowed completely unambigous assignments of atomic coordinates and water positions, yielding greater insights into GPCR activation and function. I will discuss the lessons we have learnt during this journey and show the benchmarks in both sample preparation, data collection and subsequent analysis that have given us the ability to consistently achieve high resolution cryoEM GPCR structures.