Structure Determination from Lipidic Cubic Phase Embedded Microcrystals by MicroED. Structure 2020 Oct 06;28(10):1149-1159.e4
Date
08/01/2020Pubmed ID
32735770Pubmed Central ID
PMC7544639DOI
10.1016/j.str.2020.07.006Scopus ID
2-s2.0-85089367384 (requires institutional sign-in at Scopus site) 17 CitationsAbstract
The lipidic cubic phase (LCP) technique has proved to facilitate the growth of high-quality crystals that are otherwise difficult to grow by other methods. However, the crystal size optimization process could be time and resource consuming, if it ever happens. Therefore, improved techniques for structure determination using these small crystals is an important strategy in diffraction technology development. Microcrystal electron diffraction (MicroED) is a technique that uses a cryo-transmission electron microscopy to collect electron diffraction data and determine high-resolution structures from very thin micro- and nanocrystals. In this work, we have used modified LCP and MicroED protocols to analyze crystals embedded in LCP converted by 2-methyl-2,4-pentanediol or lipase, including Proteinase K crystals grown in solution, cholesterol crystals, and human adenosine A2A receptor crystals grown in LCP. These results set the stage for the use of MicroED to analyze microcrystalline samples grown in LCP, especially for those highly challenging membrane protein targets.
Author List
Zhu L, Bu G, Jing L, Shi D, Lee MY, Gonen T, Liu W, Nannenga BLAuthors
Wei Liu PhD Associate Professor in the Pharmacology and Toxicology department at Medical College of WisconsinLan Zhu PhD Assistant Professor in the Pharmacology and Toxicology department at Medical College of Wisconsin
MESH terms used to index this publication - Major topics in bold
CholesterolCryoelectron Microscopy
Crystallization
Endopeptidase K
Glycols
Lipids
Microscopy, Electron, Transmission
Nanoparticles
Receptor, Adenosine A2A
