Extended Data Fig. 6: Comparison of two biochemical approaches to obtain GDP-bound MOR-Gαi complexes for cryoEM analysis: GDP ‘constant GDP’ versus ‘re-bound GDP’. | Nature

Extended Data Fig. 6: Comparison of two biochemical approaches to obtain GDP-bound MOR-Gαi complexes for cryoEM analysis: GDP ‘constant GDP’ versus ‘re-bound GDP’.

From: Structural snapshots capture nucleotide release at the μ-opioid receptor

Extended Data Fig. 6: Comparison of two biochemical approaches to obtain GDP-bound MOR-Gαi complexes for cryoEM analysis: GDP ‘constant GDP’ versus ‘re-bound GDP’.

a, Schematic overview of biochemical preparation with ‘constant GDP’ continuously being supplied with GDP throughout the purification process. ‘Re-bound GDP’ was treated with apyrase for complexation, followed by size-exclusion chromatography and finally supplemented with 100 µM GDP for cryoEM studies. b-c, Direct comparison of ‘constant GDP’ versus ‘re-bound GDP’ experiments for naloxone and loperamide, showing virtually identical 2D classes and 3D reconstructions of the respective state.

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