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Preparation of mini-Gs heterotrimer

Christopher Gordon Tate Christopher Gordon Tate

Last updated date: Dec 1, 2019 Views: 1704 Forks: 0

original An abbreviated version of this protocol was published in eLife in May, 2018
Cryo-EM structure of the adenosine A2A receptor coupled to an engineered heterotrimeric G protein
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The mini-GS construct (399) used in single particle cryo-EM reconstructions is based on the construct 393 that was used in the structure determination of the A2AR- mini-GS crystal structure (Carpenter et al., 2016; Carpenter and Tate, 2016). However, unlike construct 393, mini-GS399 binds βγ (Nehmé et al., 2017). The expression and purification of the respective components and assembly to make the complex containing mini-GS-β1γ2, and the preparation of nanobody Nb35, were all performed following the protocols described previously (Carpenter and Tate, 2016; Rasmussen et al., 2011; Carpenter and Tate, 2017b).

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How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Tate, C G(2019). Preparation of mini-Gs heterotrimer. Bio-protocol Preprint. bio-protocol.org/prep77.
  2. García-Nafría, J., Lee, Y., Bai, X., Carpenter, B. and Tate, C. G.(2018). Cryo-EM structure of the adenosine A2A receptor coupled to an engineered heterotrimeric G protein. eLife. DOI: 10.7554/eLife.35946

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