Key Findings:

1. IFA: This method utilizes the fluorescence energy transfer between a fluorophore and monolayer graphene oxide to measure the distance between the fluorophore and the lipid bilayer. It allows researchers to track the three-dimensional movement of the protein within the membrane and capture nanoscale changes in insertion depth.
2. LipoFRET: This technique relies on FRET between a donor fluorophore and multiple acceptor quenchers encapsulated in liposomes. By analyzing the intensity of the fluorescence, researchers can determine the penetration depth of the protein within the liposome membrane and study curvature-sensitive membrane proteins.
3. Applications: The protocol demonstrates the effectiveness of these methods by studying the insertion dynamics of MLKL and α-synuclein, two important membrane proteins involved in cell death and neurodegenerative diseases, respectively.

Significance: The development of SIFA and LipoFRET represents a significant advancement in the field of membrane protein research. These techniques offer valuable tools for understanding the structure and function of membrane proteins, and their potential applications in drug discovery and disease treatment.

The work entitled “Real-time analysis of nanoscale dynamics in membrane protein insertion via single-molecule imaging” was published on Biophysics Reports (published on December, 2024).
DOI: 10.52601/bpr.2024.240024