Optimizing the α1B-adrenergic receptor for solution NMR studies

Schuster, Matthias; Deluigi, Mattia; Pantić, Milica; Vacca, Santiago; Baumann, Christian; Scott, Daniel J; Plückthun, Andreas; Zerbe, Oliver

doi:10.1016/j.bbamem.2020.183354

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Schuster, M., Deluigi, M., Pantić, M., Vacca, S., Baumann, C., Scott, D. J., Plückthun, A., & Zerbe, O. (2020). Optimizing the α1B-adrenergic receptor for solution NMR studies. Biochimica et Biophysica Acta. Biomembranes, 1862(10), 183354. https://doi.org/10.1016/j.bbamem.2020.183354

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 de