TL;DR
The current protocol outlines a method for membrane purification utilizing ultracentrifugation and detergents, which facilitates the isolation of membrane proteins in their native conformational state.
Together, the methods described here demonstrate that careful selection via thorough screening, and optimization of expression and purification conditions, are an invaluable investment of time, ultimately yielding the best target not only for successful high-resolution structural analyses-X-ray crystallography and cryo...
This chapter outlines a detailed workflow for the expression, purification, and structural analysis of the ABC transporter MsbA from Pseudomonas aeruginosa (PaMsbA), which exemplifies how biochemical optimization, native mass spectrometry, and cryo-electron microscopy (cryo-EM) can be integrated to investigate membrane...
Transmembrane signalling by membrane proteins is essential for physiological processes and disease states, making these proteins key targets for drug development. Structural biology provides unique insights into the mechanisms of membrane protein function and dysfunction caused by mutations. Most high-resolution membra...
On-bead Reconstitution into Bilayers via Cell-free Transcription and Translation (ORB-TXTL), a platform that uses compositionally tunable lipid bilayer-coated silica beads as scaffolds for cell-free synthesized interacting and integral membrane proteins, is developed.
A generalizable workflow for producing soluble receptor domains from membrane proteins is established and validated HasR constructs suitable for downstream ligand-screening applications, including aptamer and nanobody discovery.
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