9CHV

cryo-EM structure of calcineurin-fused beta2 adrenergic receptor in apo state

Summary for 9CHV

• Entry DOI: 10.2210/pdb9chv/pdb
• Related: 9CHU
• EMDB information: 45602 45603
• Descriptor: Beta-2 adrenergic receptor,Calcineurin subunit B type 1, Protein phosphatase 3 catalytic subunit alpha, Peptidyl-prolyl cis-trans isomerase FKBP1A, ... (4 entities in total)
• Functional Keywords: gpcr, cryo-em, calcineurin fusion, inactive state, membrane protein, membrane protein-hydrolase-isomerase complex, membrane protein/hydrolase/isomerase
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 3
• Total formula weight: 106709.22

Authors

Xu, J.,Chen, G.,Du, Y.,Kobilka, B.K. (deposition date: 2024-07-02, release date: 2024-11-13, Last modification date: 2024-11-27)

Primary citation

Xu, J.,Chen, G.,Wang, H.,Cao, S.,Heng, J.,Deupi, X.,Du, Y.,Kobilka, B.K.
Calcineurin-fusion facilitates cryo-EM structure determination of a Family A GPCR.
Proc.Natl.Acad.Sci.USA, 121:e2414544121-e2414544121, 2024

PubMed Abstract: Advances in singe-particle cryo-electron microscopy (cryo-EM) have made it possible to solve the structures of numerous Family A and Family B G protein-coupled receptors (GPCRs) in complex with G proteins and arrestins, as well as several Family C GPCRs. Determination of these structures has been facilitated by the presence of large extramembrane components (such as G protein, arrestin, or Venus flytrap domains) in these complexes that aid in particle alignment during the processing of the cryo-EM data. In contrast, determination of the inactive state structure of Family A GPCRs is more challenging due to the relatively small size of the seven transmembrane domain (7TM) and to the surrounding detergent micelle that, in the absence of other features, make particle alignment impossible. Here, we describe an alternative protein engineering strategy where the heterodimeric protein calcineurin is fused to a GPCR by three points of attachment, the cytoplasmic ends of TM5, TM6, and TM7. This three-point attachment provides a more rigid link with the GPCR transmembrane domain that facilitates particle alignment during data processing, allowing us to determine the structures of the β adrenergic receptor (βAR) in the apo, antagonist-bound, and agonist-bound states. We expect that this fusion strategy may have broad application in cryo-EM structural determination of other Family A GPCRs.

PubMed: 39565314
DOI: 10.1073/pnas.2414544121

Experimental method

ELECTRON MICROSCOPY (3.95 Å)