6LPJ

A2AR crystallized in EROCOC17+4, LCP-SFX at 277 K

6LPJ の概要

• エントリーDOI: 10.2210/pdb6lpj/pdb
• 分子名称: Adenosine receptor A2a,Soluble cytochrome b562,Adenosine receptor A2a, N-OCTANE, UNDECANE, ... (14 entities in total)
• 機能のキーワード: lcp crystallization, isoprenoid-chained-lipid, gpcr, sfx, membrane protein
• 由来する生物種: Homo sapiens (Human); 詳細
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 56057.15

構造登録者

Ihara, K.,Hato, M.,Nakane, T.,Yamashita, K.,Kimura-Someya, T.,Hosaka, T.,Ishizuka-Katsura, Y.,Tanaka, R.,Tanaka, T.,Sugahara, M.,Hirata, K.,Yamamoto, M.,Nureki, O.,Tono, K.,Nango, E.,Iwata, S.,Shirouzu, M. (登録日: 2020-01-10, 公開日: 2020-11-25, 最終更新日: 2024-10-23)

主引用文献

Ihara, K.,Hato, M.,Nakane, T.,Yamashita, K.,Kimura-Someya, T.,Hosaka, T.,Ishizuka-Katsura, Y.,Tanaka, R.,Tanaka, T.,Sugahara, M.,Hirata, K.,Yamamoto, M.,Nureki, O.,Tono, K.,Nango, E.,Iwata, S.,Shirouzu, M.
Isoprenoid-chained lipid EROCOC 17+4 : a new matrix for membrane protein crystallization and a crystal delivery medium in serial femtosecond crystallography.
Sci Rep, 10:19305-19305, 2020

PubMed Abstract: In meso crystallization of membrane proteins relies on the use of lipids capable of forming a lipidic cubic phase (LCP). However, almost all previous crystallization trials have used monoacylglycerols, with 1-(cis-9-octadecanoyl)-rac-glycerol (MO) being the most widely used lipid. We now report that EROCOC mixed with 10% (w/w) cholesterol (Fig. 1) serves as a new matrix for crystallization and a crystal delivery medium in the serial femtosecond crystallography of Adenosine A receptor (AR). The structures of EROCOC-matrix grown AR crystals were determined at 2.0 Å resolution by serial synchrotron rotation crystallography at a cryogenic temperature, and at 1.8 Å by LCP-serial femtosecond crystallography, using an X-ray free-electron laser at 4 and 20 °C sample temperatures, and are comparable to the structure of the MO-matrix grown AR crystal (PDB ID: 4EIY). Moreover, X-ray scattering measurements indicated that the EROCOC/water system did not form the crystalline L phase at least down to - 20 °C, in marked contrast to the equilibrium MO/water system, which transforms into the crystalline L phase below about 17 °C. As the L phase formation within the LCP-matrix causes difficulties in protein crystallography experiments in meso, this feature of EROCOC will expand the utility of the in meso method.

PubMed: 33168855
DOI: 10.1038/s41598-020-76277-x

実験手法

X-RAY DIFFRACTION (1.8 Å)