Determining the three-dimensional structure of membrane proteins has historically been a huge challenge for scientists due to the difficulties in expression, solubilization, and purification of these proteins. Recent advancements in membrane protein sample preparation have allowed researchers to overcome many of these challenges, facilitating cryo-EM structure determination.

 

In this webinar, Dr. Mahfuz Rahman will present the overall workflow of single-particle cryo-EM sample preparation. He will discuss the current challenges of membrane protein sample preparation and share strategies and best practices to improve the success rate of sample preparation and obtain high-quality cryo-EM results. In addition, Dr. Rosemary Cater will discuss an approach used to solve structures of small membrane proteins using cryo-EM: complexation of target proteins with specific monoclonal antigen-binding fragments (Fab) to increase particle size and provide fiducials outside of the transmembrane region for alignment. She will describe how her lab used this approach to determine the cryo-EM structure of Major Facilitator Superfamily Domain containing 2A (MFSD2A; molecular weight of 58 kDa) to a resolution of 3.0 Å.

 

Learning Objectives