Dark nanodiscs for evaluating membrane protein thermostability by differential scanning fluorimetry.

ABSTRACT: Measuring protein thermostability provides valuable information on the biophysical rules that govern the structure-energy relationships of proteins. However, such measurements remain a challenge for membrane proteins. Here, we introduce a new experimental system to evaluate membrane protein thermostability. This system leverages a recently developed nonfluorescent membrane scaffold protein to reconstitute proteins into nanodiscs and is coupled with a nano-format of differential scanning fluorimetry (nanoDSF). This approach offers a label-free and direct measurement of the intrinsic tryptophan fluorescence of the membrane protein as it unfolds in solution without signal interference from the "dark" nanodisc. In this work, we demonstrate the application of this method using the disulfide bon

SUBMITTER: Selvasingh JA 

PROVIDER: S-EPMC10808023 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature