Probing Membrane Protein Assembly into Nanodiscs by In Situ Dynamic Light Scattering: A2A Receptor as a Case Study

Biology
Rosana I Reis, Isabel Moraes

Abstract

Membrane proteins play a crucial role in cell physiology by participating in a variety of essential processes such as transport, signal transduction and cell communication. Hence, understanding their structure-function relationship is vital for the improvement of therapeutic treatments. Over the last decade, based on the development of detergents, amphipoles and styrene maleic-acid lipid particles (SMALPs), remarkable accomplishments have been made in the field of membrane protein structural biology. Nevertheless, there are still many drawbacks associated with protein-detergent complexes, depending on the protein in study or experimental application. Recently, newly developed membrane mimetic systems have become very popular for allowing a structural and functional characterisation of membrane proteins in vitro. The nanodisc technology is one such valuable tool, which provides a more native-like membrane environment than detergent micelles or liposomes. In addition, it is also compatible with many biophysical and biochemical methods. Here we describe the use of in situ dynamic light scattering to accurately and rapidly probe membrane proteins' reconstitution into nanodiscs. The adenosine type 2A receptor (A2AR) was used as a ca...Continue Reading

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Citations

Nov 20, 2020·Biology·James BirchAndrew Quigley

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Methods Mentioned

BETA
X-ray
nuclear magnetic resonance
protein folding
electron
NMR
surface plasmon resonance
Dynamic Light Scattering
electrophoresis
size exclusion chromatography

Software Mentioned

situ DLS
in
DLS SpectroLight

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