Journal article

Optimizing the alpha(1B)-adrenergic receptor for solution NMR studies

Matthias Schuster, Mattia Deluigi, Milica Pantic, Santiago Vacca, Christian Baumann, Daniel J Scott, Andreas Pluckthun, Oliver Zerbe

BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES | ELSEVIER | Published : 2020

Abstract

Sample preparation for NMR studies of G protein-coupled receptors faces special requirements: Proteins need to be stable for prolonged measurements at elevated temperatures, they should ideally be uniformly labeled with the stable isotopes 13C, 15N, and all carbon-bound protons should be replaced by deuterons. In addition, certain NMR experiments require protonated methyl groups in the presence of a perdeuterated background. All these requirements are most easily satisfied when using Escherichia coli as the expression host. Here we describe a workflow, starting from a temperature-stabilized mutant of the α1B-adrenergic receptor, obtained using the CHESS methodology, into an even more stable ..

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Grants

Awarded by Swiss National Science Foundation


Awarded by Research Foundation of the University of Zurich


Funding Acknowledgements

This project is supported by a grant from the Swiss National Science Foundation (grant no. 310030_179314 to O.Z.) and from the Research Foundation of the University of Zurich (grant no. FK-18-083 to C.B.). We deeply thank Franz Hagn for a gift of the MSP Delta H5 expression plasmid.