Enhanced Site-Specific Fluorescent Labeling of Membrane Proteins Using Native Nanodiscs.

ABSTRACT: Fluorescent labeling of membrane proteins is essential for exploring their functions, signaling pathways, interaction partners, and structural dynamics. Organic fluorophores are commonly used for this purpose due to their favorable photophysical properties and photostability. However, a persistent challenge is the inaccessibility of the surface-exposed cysteine residues required for site-specific labeling, as these residues often become sequestered within detergent micelles during protein extraction. To address this limitation, we developed an approach based on polymer-encapsulated nanodiscs that preserves the protein's native-like lipid-bilayer environment while ensuring the accessibility of surface-exposed cysteine residues. In this method, His-tagged proteins embedded in native nanodisc

SUBMITTER: Ezsias B 

PROVIDER: S-EPMC11852578 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature