LINXS Event - Membrane Protein workshop: Structural Resolution of Membrane Proteins: From Expression to Sample Preparation

Session

Session 3 - Chair: Maria Gourdon

25 May 2021, 15:30

30. Keynote talk - Preparation of lipid flippases for cryo-EM studies

Joseph Lyons (Aarhus University, Denmark)

25/05/2021, 15:30

P4-ATPases are lipid flippases that drive the ATP-dependent inward translocation (flipping) of lipids within the membrane. They are members of the P-type ATPase superfamily and largely function as binary complexes with an auxiliary protein from the CDC50 family. Select lipid flippases are auto-regulated by conserved motifs in their termini and require activation by an external stimulus, i.e.,...

31. Contr. talk - Production and purification of the K+-dependent P-Type ATPase KdpFABC for crystallization and cryo-EM

Charlott Stock (Aarhus University)

25/05/2021, 16:10

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...