7RM5

MicroED structure of the human adenosine receptor at 2.8A

Summary for 7RM5

• Entry DOI: 10.2210/pdb7rm5/pdb
• EMDB information: 24551
• Descriptor: Adenosine receptor A2a/Soluble cytochrome b562 chimera, 4-{2-[(7-amino-2-furan-2-yl[1,2,4]triazolo[1,5-a][1,3,5]triazin-5-yl)amino]ethyl}phenol, CHOLESTEROL, ... (5 entities in total)
• Functional Keywords: membrane protein
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 1
• Total formula weight: 51881.22

Authors

Martynowycz, M.W.,Shiriaeva, A.,Ge, X.,Hattne, J.,Nannenga, B.L.,Cherezov, V.,Gonen, T. (deposition date: 2021-07-26, release date: 2021-09-08, Last modification date: 2024-11-20)

Primary citation

Martynowycz, M.W.,Shiriaeva, A.,Ge, X.,Hattne, J.,Nannenga, B.L.,Cherezov, V.,Gonen, T.
MicroED structure of the human adenosine receptor determined from a single nanocrystal in LCP.
Proc.Natl.Acad.Sci.USA, 118:-, 2021

PubMed Abstract: G protein-coupled receptors (GPCRs), or seven-transmembrane receptors, are a superfamily of membrane proteins that are critically important to physiological processes in the human body. Determining high-resolution structures of GPCRs without bound cognate signaling partners, such as a G protein, requires crystallization in lipidic cubic phase (LCP). GPCR crystals grown in LCP are often too small for traditional X-ray crystallography. These microcrystals are ideal for investigation by microcrystal electron diffraction (MicroED), but the gel-like nature of LCP makes traditional approaches to MicroED sample preparation insurmountable. Here, we show that the structure of a human A adenosine receptor can be determined by MicroED after converting the LCP into the sponge phase followed by focused ion-beam milling. We determined the structure of the A adenosine receptor to 2.8-Å resolution and resolved an antagonist in its orthosteric ligand-binding site, as well as four cholesterol molecules bound around the receptor. This study lays the groundwork for future structural studies of lipid-embedded membrane proteins by MicroED using single microcrystals that would be impossible with other crystallographic methods.

PubMed: 34462357
DOI: 10.1073/pnas.2106041118

Experimental method

ELECTRON CRYSTALLOGRAPHY (2.79 Å)