Abstract
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.
Original language | English (US) |
---|---|
Pages (from-to) | 1149-1159.e4 |
Journal | Structure |
Volume | 28 |
Issue number | 10 |
DOIs | |
State | Published - Oct 6 2020 |
Keywords
- GPCR
- Proteinase K
- additive phase conversion
- cholesterol
- cryo-electron microscopy (cryo-EM)
- lipase hydrolysis
- lipidic cubic phase (LCP)
- membrane protein
- microcrystal electron diffraction (MicroED)
- microcrystallography
ASJC Scopus subject areas
- Structural Biology
- Molecular Biology
Fingerprint
Dive into the research topics of 'Structure Determination from Lipidic Cubic Phase Embedded Microcrystals by MicroED'. Together they form a unique fingerprint.Datasets
-
LCP-embedded Proteinase K treated with MPD
Zhu, L. (Contributor), Bu, G. (Contributor), Jing, L. (Contributor), Shi, D. (Contributor), Lee, M. (Contributor), Gonen, T. (Contributor), Liu, W. (Contributor) & Nannenga, B. (Contributor), Protein Data Bank (PDB), Aug 5 2020
DOI: 10.2210/pdb6PQ0, https://www.wwpdb.org/pdb?id=pdb_00006pq0
Dataset
-
LCP-embedded Proteinase K treated with lipase
Zhu, L. (Contributor), Bu, G. (Contributor), Jing, L. (Contributor), Shi, D. (Contributor), Lee, M. (Contributor), Gonen, T. (Contributor), Liu, W. (Contributor) & Nannenga, B. (Contributor), Protein Data Bank (PDB), Aug 5 2020
DOI: 10.2210/pdb6PQ4, https://www.wwpdb.org/pdb?id=pdb_00006pq4
Dataset