8TZQ
CryoEM structure of D2 dopamine receptor in complex with GoA KE Mutant, scFv16, and dopamine
Summary for 8TZQ
| Entry DOI | 10.2210/pdb8tzq/pdb |
| EMDB information | 41766 |
| Descriptor | scFv16, Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1, Guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2, ... (6 entities in total) |
| Functional Keywords | gpcr, dopamine, drd2, dominant negative, membrane protein |
| Biological source | Escherichia coli More |
| Total number of polymer chains | 5 |
| Total formula weight | 166887.12 |
| Authors | Krumm, B.E.,Kapolka, N.J.,Fay, J.F.,Roth, B.L. (deposition date: 2023-08-27, release date: 2024-08-21, Last modification date: 2025-05-14) |
| Primary citation | Knight, K.M.,Krumm, B.E.,Kapolka, N.J.,Ludlam, W.G.,Cui, M.,Mani, S.,Prytkova, I.,Obarow, E.G.,Lefevre, T.J.,Wei, W.,Ma, N.,Huang, X.P.,Fay, J.F.,Vaidehi, N.,Smrcka, A.V.,Slesinger, P.A.,Logothetis, D.E.,Martemyanov, K.A.,Roth, B.L.,Dohlman, H.G. A neurodevelopmental disorder mutation locks G proteins in the transitory pre-activated state. Nat Commun, 15:6643-6643, 2024 Cited by PubMed Abstract: Many neurotransmitter receptors activate G proteins through exchange of GDP for GTP. The intermediate nucleotide-free state has eluded characterization, due largely to its inherent instability. Here we characterize a G protein variant associated with a rare neurological disorder in humans. Gα has a charge reversal that clashes with the phosphate groups of GDP and GTP. As anticipated, the purified protein binds poorly to guanine nucleotides yet retains wild-type affinity for G protein βγ subunits. In cells with physiological concentrations of nucleotide, Gα forms a stable complex with receptors and Gβγ, impeding effector activation. Further, we demonstrate that the mutant can be easily purified in complex with dopamine-bound D2 receptors, and use cryo-electron microscopy to determine the structure, including both domains of Gα, without nucleotide or stabilizing nanobodies. These findings reveal the molecular basis for the first committed step of G protein activation, establish a mechanistic basis for a neurological disorder, provide a simplified strategy to determine receptor-G protein structures, and a method to detect high affinity agonist binding in cells. PubMed: 39103320DOI: 10.1038/s41467-024-50964-z PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (3.2 Å) |
Structure validation
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