Addressing the challenges of membrane protein sample preparation for cryo-EM analysis

Presented By:
Mahfuz Rahman
Rosemary Cater

Speaker Biography:
Mahfuz Rahman is an R&D Staff Scientist at Thermo Fisher Scientific in Rockford, IL. He graduated from the University of Dhaka, Bangladesh, with a major in pharmacy. Following graduation, he worked as a product manager in the pharmaceutical industry for several years before entering graduate school. During his doctoral research, he pursued the structural analysis of ABC transporters under the supervision of Dr. Yoshitsugu Shiro at the RIKEN SPring-8 Center in Japan. As a postdoctoral fellow, he studied the molecular mechanism of ligand-gated ion channels with Dr. Ryan Hibbs at UT Southwestern Medical Center in Dallas, TX. His expertise includes molecular biology, protein biochemistry, spectroscopy, pharmacology, X-ray crystallography (Ph.D.), single-particle cryo-EM (postdoc), and membrane proteins. Dr. Rahman joined Thermo Fisher Scientific in 2022, targeting method/reagent development for protein expression, extraction, and purification to facilitate the structure determination of challenging and complex membrane proteins.

Rosemary Cater is an Associate Research Scientist at Columbia University, New York. She leverages her expertise in structural biology, membrane protein biochemistry, and biophysics to understand some of the brain’s most elusive yet important proteins. Dr. Cater fulfilled her PhD at the University of Sydney under the supervision of Professor Renae Ryan and Professor Robert Vandenberg, where she used electrophysiology and X-ray crystallography to understand how glutamate transporters serve a secondary role as chloride channels. Since 2017, Dr. Cater has worked in the laboratory of Professor Filippo Mancia, where she uses single-particle cryo-EM and antigen-binding technology to determine structures of small membrane proteins. The overarching goal of her research is to understand the molecular mechanisms of transport at the blood-brain barrier, and her current research focuses on the transport of omega-3 fatty acids into the brain.

Webinar:
Addressing the challenges of membrane protein sample preparation for cryo-EM analysis

Webinar Abstract:
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

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