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DOI	10.1073/pnas.2106041118
	MicroED structure of the human adenosine receptor determined from a single nanocrystal in LCP
	Martynowycz M.W.; Shiriaeva A.; Ge X.; Hattne J.; Nannenga B.L.; Cherezov V.; Gonen T.
发表日期	2021
ISSN	0027-8424
卷号	118 期号:36
英文摘要	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 A2A 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 A2A 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. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	GPCR; Ion-beam milling; Lipidic cubic phase; Membrane proteins; MicroED
语种	英语
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/238404
作者单位	HHMI, University of California, Los Angeles, CA 90095, United States; Department of Biological Chemistry, University of California, Los Angeles, CA 90095, United States; Bridge Institute, Michelson Center for Convergent Biosciences, University of Southern California, Los Angeles, CA 90007, United States; Department of Chemistry, University of Southern California, Los Angeles, CA 90007, United States; Mork Family Department of Chemical Engineering and Materials Science, University of Southern California, Los Angeles, CA 90007, United States; Chemical Engineering, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, AZ 85287, United States; Biodesign Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, AZ 85287, United States; Department of Physiology, University of California, Los Angeles, CA 90095, United States
推荐引用方式 GB/T 7714	Martynowycz M.W.,Shiriaeva A.,Ge X.,et al. MicroED structure of the human adenosine receptor determined from a single nanocrystal in LCP[J],2021,118(36).
APA	Martynowycz M.W..,Shiriaeva A..,Ge X..,Hattne J..,Nannenga B.L..,...&Gonen T..(2021).MicroED structure of the human adenosine receptor determined from a single nanocrystal in LCP.Proceedings of the National Academy of Sciences of the United States of America,118(36).
MLA	Martynowycz M.W.,et al."MicroED structure of the human adenosine receptor determined from a single nanocrystal in LCP".Proceedings of the National Academy of Sciences of the United States of America 118.36(2021).