Illuminating the energy landscape of GPCRs: the key contribution of solution-state NMR associated with Escherichia coli as an expression host.
Related Articles Illuminating the energy landscape of GPCRs: the key contribution of solution-state NMR associated with Escherichia coli as an expression host.
Biochemistry. 2018 Apr 02;:
Authors: Casiraghi M, Damian M, Lescop E, Baneres JL, Catoire LJ
Abstract
Conformational dynamics of GPCRs are central to their function but are difficult to explore at the atomic scale. Solution-state NMR has provided the major contribution in that area during the past decade, despite non-optimized labeling schemes due to the use of insect cells, and to a lesser extent yeast, as the main expression hosts. Indeed, the most efficient isotope-labeling scheme ever to address energy landscape issues for large proteins or protein complexes relies on the use of 13CH3 probes immersed in a perdeuterated dipolar environment, which is essentially out of reach of eukaryotic expression systems. In contrast, although its contribution has been underestimated because of technical issues, Escherichia coli is by far the best-adapted host for such labeling. As it is now tightly controlled, we show in this review that bacterial expression can provide a NMR spectral resolution never achieved in the GPCR field.
PMID: 29607648 [PubMed - as supplied by publisher]
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