12.05.2020: Webinar announcement (May 19): Insights and New Approaches in Transporter Characterization
On the 19th of May we continue our Webinar-Series with a talk from Prof. Dr. Camilo Perez, Nathan Thomas and Dr. Maria Barthmes:
"INSIGHTS AND NEW APPROACHES IN TRANSPORTER CHARACTERIZATION - SURFE²R N1"
Date: 19. May 2020
Time: 4:00 PM CET / 10:00 AM EDT
Prof. Dr. Camilo Perez (Head of Research Group - BIOZENTRUM, University of Basel, Switzerland)
Nathan Thomas (5th Year PhD, University of Wisconsin-Madison [Dr. Henzler-Wildman's lab])
Dr. Maria Barthmes (Product Manager, Nanion Technologies)
"Maria will introduce SSM-based Electrophysiology going over basic features and principles of the method and have a look at experimental workflows.
SSM-based electrophysiology is a capacitive sensor-based method to detect membrane currents generated by low turnover proteins such as transporters and membrane pumps. By resolving protein activity in real time and label free it introduces the advantages of electrophysiology to the field of membrane transporters.
"Unlocking the (Reversal) Potential of SSM Electrophysiology: Transporter Stoichiometry with the SURFE²R N1”
Mechanistic studies of ion-coupled transporters can be facilitated if the stoichiometry of the transport reaction is known, but current methods for measuring transport stoichiometry are both material and labor intensive. We developed an SSM-based electrophysiology reversal potential assay using the SURFE²R N1. This assay allows for fast, accurate determination of transport stoichiometry without the need for radioactivity or fluorescent probes while using only picomols of protein. We report our results with GdX, aguanidinium exchanger with a known 2:1 proton:guanidinium antiport stoichiometry.“
"Characterization of a choline uniporter by SSM-based electrophysiology"
My group studies multiple transport proteins involved in the biosynthesis of teichoic acids in Gram-positive bacteria. We aim to understand the mechanism of these proteins and to describe their importance for bacterial growth, adaptation, and survival, as they are considered valuable drug targets. Some Gram-positive bacteria display teichoic acids with a phosphorylcholine modification that serves as an anchor for choline-binding proteins (CBP). CBPs play fundamental roles in interactions with biological surfaces and in response to external threats. The growth of these bacteria frequently depends onexogenous choline, which is metabolized to modify teichoic acids. Transporters located at their plasma membrane harvest large amounts of choline from the external medium. We are using SSM-based electrophysiology to characterize the function of some of these transporters. SSM-based electrophysiology allows us to perform high-throughput screening of multiple chemical species, providing the biochemical basis for the production of novel inhibitors of Gram-positive bacteria choline transporters.