Detergent-free extraction of membrane proteins and lipids from native cells
EMSL Project ID
60481
Abstract
Membrane proteins account for >30% of most genomes, yet only make up <4% of reported high-resolution structures in the protein data bank. Current approaches for isolating membrane proteins from native cells are tedious, labor intensive, and require "mild" detergents that alter the native protein structure and lipid environment. The objective of this project is to engineer a library of synthetic “scaffolds proteins” that bypass detergent solubilization procedures to directly extract membrane protein-lipid complexes into affinity purifiable nanodiscs. To achieve this objective, we will fuse anti-microbial peptides (AMPs) to commercially available membrane scaffold proteins (MSP). AMPs contain the inherent ability to naturally disrupt and “dig” out cell membrane constituents. By conferring these detergent-like qualities of AMPs to the MSP scaffold, we will generate a reagent capable of directly extract membrane proteins and surrounding lipids into nanodiscs for structure-function analysis. We will use the MSP-AMP library to capture membrane proteins and surrounding lipids from two model organisms important for biofuel production: the algae Ostreococcus tauri and the yeast Yarrowia lipolytica. Upon successful extraction and demonstration of proof-of-principle, we will make the library available to the EMSL user community.
Project Details
Start Date
2022-10-01
End Date
N/A
Status
Active
Released Data Link
Team
Principal Investigator
Team Members