Work

Native Mass Spectrometry of Membrane Protein Lipid and Ligand Complexes Using de novo Designed WRAP Domains

Year: 2026

Type: article

Abstract: Membrane proteins engage in dynamic transient interactions with surrounding lipids along with their substrates. Capturing these concomitant contacts often remains a challenging problem. Here, we demonstrate that generative deep learning-designed WRAP domains can preserve weak interactions of membrane proteins in native mass spectrometry. Using WRAP-fused GlpG, AqpM, and OmpA as model systems, we show that lipids can bind in multiple positions inside the WRAP, allowing for their characterization by native mass spectrometry. We show that lipid interactions can be altered by changing the WRAP structure. We use the approach to show that WRAP–OmpA binds phosphatidylethanolamine lipids within cavities formed at the protein–WRAP interface, while simultaneously accommodating weak chitobiose ligand binding. These findings establish WRAPs as versatile vehicles to probe ligand and lipid interactions of membrane proteins.

Source: Journal of the American Chemical Society

DOI: 10.1021/jacs.6c15293