VIDEO - Membrane Proteins Workshop - Production and purification of the K+-dependent P-Type ATPase KdpFABC for crystallization and cryo-EM with Charlott Stack
VIDEO - Membrane Proteins Workshop - Production and purification of the K+-dependent P-Type ATPase KdpFABC for crystallization and cryo-EM with Charlott Stack
Speaker: Charlott Stack, Aarhus University, Denmark
This lecture was part of the workshop, Structural Resolution of Membrane Proteins: From Expression to Sample , organised in May 2021 by the LINXS working group Membrane Proteins, which fall under the Integrative Structural Biology theme.
Abstract
At very low extracellular K+ concentrations (<100 μM), KdpFABC rescues bacterial cells by taking up K+ against an up to 105-fold concentration gradient. To understand the mechanism of this heterotetrameric complex, KdpFABC was purified for functional and structural investigations. A new expression system for KdpFABC was generated, a 3-step chromatographic purification implemented and protein functionality in different detergents as well as in the presence of inhibitors was tested. The work was performed in the lab of Prof. Inga Hänelt at the University of Frankfurt from 2014-2020.