Journal
STRUCTURE
Volume 28, Issue 10, Pages 1149-+Publisher
CELL PRESS
DOI: 10.1016/j.str.2020.07.006
Keywords
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Funding
- NSF MRI [1531991]
- Center for Applied Structural Discovery (CASD) at the Biodesign Institute at Arizona State University
- Mayo Clinic ASU Collaborative Seed Grant Award
- Flinn Foundation Seed Grant
- STC Program of the National Science Foundation through BioXFEL [1231306]
- National Institutes of Health [R21DA042298, R01GM124152, P41GM136508]
- Howard Hughes Medical Institute
- Div Of Biological Infrastructure
- Direct For Biological Sciences [1531991, 1231306] Funding Source: National Science Foundation
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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 A(2A) 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.
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