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-Structure paper
Title | MicroED structure of the human adenosine receptor determined from a single nanocrystal in LCP. |
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Journal, issue, pages | Proc Natl Acad Sci U S A, Vol. 118, Issue 36, Year 2021 |
Publish date | Sep 7, 2021 |
Authors | Michael W Martynowycz / Anna Shiriaeva / Xuanrui Ge / Johan Hattne / Brent L Nannenga / Vadim Cherezov / Tamir Gonen / |
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 ...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. |
External links | Proc Natl Acad Sci U S A / PubMed:34462357 / PubMed Central |
Methods | EM (electron crystallography) |
Resolution | 2.79 Å |
Structure data | EMDB-24551, PDB-7rm5: |
Chemicals | ChemComp-ZMA: ChemComp-CLR: ChemComp-NA: ChemComp-HOH: |
Source |
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Keywords | MEMBRANE PROTEIN |