Nanodisc-Tm: Rapid functional assessment of nanodisc reconstituted membrane proteins by CPM assay

摘要

class="kwd-title">Method name: Nanodisc-Tm class="kwd-title">Keywords: Membrane proteins, Nanodiscs, CPM assay class="head no_bottom_margin" id="abs0010title">AbstractMembrane proteins are generally unstable in detergents. Therefore, biochemical and biophysical studies of membrane proteins in lipidic environments provides a near native-like environment suitable for membrane proteins. However, manipulation of proteins embedded in lipid bilayer has remained difficult. Methods such as nanodiscs and lipid cubic phase have been developed for easy manipulation of membrane proteins and have yielded significant insights into membrane proteins. Traditionally functional reconstitution of receptors in nanodiscs has been studied with radioligands. We present a simple and faster method for studying the functionality of reconstituted membrane proteins for routine characterization of protein batches after initial optimization of suitable conditions using radioligands. The benefits of the method are class="first-line-outdent">

• Faster and generic method to assess functional reconstitution of membrane proteins.

• Adaptable in high throughput format (≥96 well format).

• Stability measurement in near-native lipid environment and lipid dependent melting temperatures.