Circularized fluorescent nanodiscs for probing protein-lipid interactions

Area of Science:

• Membrane Biology
• Biochemistry
• Biophysics

Background:

• Protein-lipid interactions are crucial for transmembrane signaling pathways.
• Existing tools for studying these interactions are limited, hindering research into signal transduction.
• Advanced methods are needed to characterize membrane biochemical reactions.

Purpose of the Study:

• To engineer a novel nanodisc-based fluorescent sensor for reporting membrane biochemical reactions.
• To create a versatile tool for detecting membrane binding and remodeling events.
• To facilitate the study of protein-lipid interactions in biological systems.

Main Methods:

• Engineered nanodiscs by circularizing them via split GFP.
• Developed an intensity-based fluorescent sensor (isenND).
• Tested isenND's specificity and robustness with various membrane-interacting proteins and peptides.

Main Results:

• The isenND sensor demonstrated robust and specific responses to diverse membrane-interacting proteins and peptides.
• The sensor successfully detected binding and remodeling events mediated by proteins like synaptotagmin and synuclein.
• Viral fusion peptides from HIV-1 and SARS-CoV-2 also elicited a response from isenND.

Conclusions:

• The engineered nanodisc-based fluorescent sensor (isenND) is a powerful tool for studying membrane biochemical reactions.
• isenND offers a versatile approach for basic and translational research in membrane biology.
• This innovation addresses the scarcity of simple tools for characterizing vital protein-lipid interactions.